LOCUS Exported 6482 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses the genetically-encoded fluorescent norepinephrine(NE) sensor GRAB_NE1h in neurons. ACCESSION . VERSION . KEYWORDS pAAV-hSyn-GRAB_NE1h 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 46..175 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" regulatory 626..635 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" sig_peptide 632..694 /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader protein_bind 695..719 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind 1822..1843 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1949..1970) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2210..2229) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 2654..3242 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2707..2727) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3125..3136) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3274..3750 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3473..3492) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3790..3930 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3790..3919 /label=AAV2 ITR rep_origin 4005..4460 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4092..4111) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4302..4323 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4477..4496) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4596..4618 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4656..4674) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4742..4846 /gene="bla" /label=AmpR promoter CDS 4847..5707 /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(5065..5084) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5878..6466 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6367..6386 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc 61 gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca 121 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc 181 cgcacgcgta tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac 241 aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac 301 aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc 361 ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc 421 ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca 481 gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc 541 ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc 601 ctgagagcgc agtcgagaaa ccggtgccac catggagaca gacacactcc tgctatgggt 661 actgctgctc tgggttccag gttccactgg tgacacaagt ttgtacaaaa aagttggcac 721 caccggtggc tccctgcagc cggacgcggg caacgcgagc tggaacggga ccgaggcgcc 781 ggggggcggc gcccgggcca ccccttactc cctgcaggtg acgctgacgc tggtgtgcct 841 ggccggcctg ctcatgctgc tcaccgtgtt cggcaacgtg ctcgtcatca tcgccgtgtt 901 cacgagccgc gcgctcaagg cgccccaaaa cctcttcctg gtgtctctgg cctcggccga 961 catcctggtg gccacgctcg tcatcccttt ctcgctggcc aacgaggtca tgggctactg 1021 gtacttcggc aaggcttggt gcgagatcta cctggcgctc gacgtgctct tctgcacgtc 1081 gtccatcgtg cacctgtgcg ccatcagcct ggaccgctac tggtccatca cacaggccat 1141 cgagtacaac ctgaagcgca cgccgcgccg catcaaggcc atcatcatca ccgtgtgggt 1201 catctcggcc gtcatctcct tcccgccgct catctccatc gagaagaagg gcggcggcgg 1261 cggcccgcag ccggccgagc cgcgctgcga gatcaacgac cagaagtggt acgtcatctc 1321 gtcgtgcatc ggctccttct tcgctccctg cctcatcatg atcctggtct acgtgcgcat 1381 ctaccagatc gccaagcgtc gcacccgcgt gccacccagc cgccggggtc cggacgccgt 1441 cgccgcgccg ccggggggca ccgagcgcag gcccaacggt ctgggccccg agcgcagcgc 1501 gggcccgggg ggcgcagagg ccgaaccgct gcccacccag ctcaacggcg cccctggcga 1561 gcccgcgccg gccgggccgc gcgacaccga cgcgctggac ctggaggagg gaggaaacgt 1621 ctatatcaag gccgacaagc agaagaacgg catcaaggcg aacttccaca tccgccacaa 1681 catcgaggac ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga 1741 cggccccgtg ctgctgcccg acaaccacta cctgagcgtg cagtccaaac tttcgaaaga 1801 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac 1861 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatggtga gaaagggcga 1921 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca 1981 caagttcagc gtgtccggcg agggtgaggg cgatgccacc tacggcaagc tgaccctgaa 2041 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac 2101 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa 2161 gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa 2221 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct 2281 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacac 2341 cggagcagca gcacgctggc gcgggcggca gaaccgcgag aagcgcttca agttcgtgct 2401 ggccgtggtc atcggagtgt tcgtggtgtg ctggttcccc ttcttcttca cctacacgct 2461 cacggccgtc gggtgctccg tgccacgcac gctcttcaaa ttcttcttct ggttcggcta 2521 ctgcaacagc tcgttgaacc cggtcatcta caccatcttc aaccacgatt tccgccgcgc 2581 cttcaagaag atcctctgtc ggggggacag gaagcggatc gtgttgtgag aattcgatat 2641 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct 2701 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc 2761 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct 2821 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga 2881 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc 2941 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac 3001 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt 3061 tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt 3121 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc 3181 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc 3241 gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca 3301 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa 3361 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg 3421 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg 3481 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt 3541 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg 3601 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc 3661 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag 3721 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg 3781 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc 3841 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag 3901 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc 3961 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg 4021 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 4081 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 4141 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 4201 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac 4261 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 4321 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat 4381 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa 4441 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata 4501 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct 4561 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt 4621 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata 4681 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt 4741 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag 4801 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca 4861 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc 4921 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat 4981 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc 5041 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg 5101 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc 5161 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat 5221 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga 5281 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc 5341 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc 5401 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt 5461 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc 5521 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc 5581 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca 5641 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca 5701 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt 5761 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta 5821 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg 5881 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc 5941 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag 6001 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa 6061 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc 6121 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc 6181 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta 6241 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag 6301 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct 6361 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga 6421 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc 6481 gg //