LOCUS Exported 6200 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses iAChSnFR (non-binding) under hSynapsin promoter. ACCESSION . VERSION . KEYWORDS pAAV.hSynap.iAChSnFR-NULL SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6200) TITLE A genetically encoded fluorescent sensor for in vivo acetylcholine detection in mice, fish, worms and flies. (iAChSnFR) JOURNAL Unpublished REFERENCE 2 (bases 1 to 6200) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6200 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 55..184 /label=AAV2 ITR promoter 266..713 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 742..751 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" sig_peptide 748..810 /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader primer_bind 1260..1281 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1384..1405) /label=EGFP-N /note="EGFP, reverse primer" CDS 2392..2421 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2425..2571 /codon_start=1 /product="transmembrane domain from platelet derived growth factor receptor beta" /label=PDGFR-beta TM domain /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ KKPR" misc_feature 2587..3175 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2640..2660) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3058..3069) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3178..3194) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3179..3195) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 3224..3345 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3235..3254) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3289..3308 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 3507..3636 /label=AAV2 ITR primer_bind complement(3655..3672) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3655..3671) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3664..3686) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3813..4268 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3900..3919) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4110..4131 /label=F1ori-F /note="F1 origin, forward primer" promoter 4294..4398 /gene="bla" /label=AmpR promoter CDS 4399..5259 /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(4617..4636) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5430..6018 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5919..5938 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6172..6189 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctgcgc 61 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc 121 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt 181 tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag 241 atctctgcag agggccctgc gtatgagtgc aagtgggttt taggaccagg atgaggcggg 301 gtgggggtgc ctacctgacg accgaccccg acccactgga caagcaccca acccccattc 361 cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt 421 gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc 481 accgccgcct cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg caaactcccc 541 ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg caccacgcga 601 ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc gactcagcgc 661 tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg cagtcgaatt 721 caagctgcta gcaaggatcc cgccaccatg gagacagaca cactcctgct atgggtactg 781 ctgctctggg ttccaggttc cactggtgac agatctgcga acgacaccgt agttgtgggc 841 tcgatcatct ttacagaagg gattatcgtc gcaaacatgg tggcagagat gattgaggcg 901 catacagacc ttaaggtggt tcgcaaactg aaccttggcg gggtcaacgt taactttgaa 961 gccattaaac gcggaggtgc gaataatggt attgacattt acgtggagta cactgggcac 1021 ggtcttgtgg atattctggg gttcccggag ccgaacgtct atatcaccgc cgacaagcag 1081 aagaacggca tcaaggcgaa cttcaagatc cgccacaacg tggaggacgg cagcgtgcag 1141 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac 1201 aaccactacc tgagcaccca gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac 1261 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac 1321 aagggcggta ccggagggag catgagcaag ggcgaggagc tgttcaccgg ggtggtgccc 1381 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgcg cggcgagggc 1441 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg 1501 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc 1561 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc 1621 caggagcgca ccatcagctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag 1681 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac 1741 ggcaacatcc tggggcacaa gctggagtac aactttccgc cgcccgcgac tactgatcca 1801 gaaggtgcat acgaaaccgt gaagaaggag tacaaacgta aatggaatat tgtatggctc 1861 aaaccactgg gattcaacaa tacggcgacg cttaccgtta aagacgaact ggcgaaacag 1921 tataacctta aaaccttcag tgacttagcg aaaatctcgg ataagctgat tctgggtgca 1981 acgatgttct ttttagaagg gcccgatggt tacccaggcc tgcaaaaact gtacaatttc 2041 aaattcaagc acaccaaaag catggacatg ggtattcgct ataccgccat tgataataac 2101 gaagttcagg taattgatgc ctgggccact gatggcttgc tggtgagcca caaattaaaa 2161 attctggagg atgataaagc gttcttcccg ccgtattatg ctgcccccat catccgtcag 2221 gatgtcttag ataagcatcc tgaactgaag gacgtgctga acaaactcgc gaatcaaatt 2281 tcactggaag aaatgcagaa actgaattac aaggtggacg gtgagggtca ggacccagcg 2341 aaagtagcta aggagttttt gaaagagaaa ggtttaattc tgcaggtcga cgaacaaaaa 2401 ctcatctcag aagaggatct gaatgctgtg ggccaggaca cgcaggaggt catcgtggtg 2461 ccacactcct tgccctttaa ggtggtggtg atctcagcca tcctggccct ggtggtgctc 2521 accatcatct cccttatcat cctcatcatg ctttggcaga agaagccacg ttagaagctt 2581 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat 2641 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct 2701 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag 2761 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc 2821 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc 2881 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct 2941 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg 3001 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg 3061 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg 3121 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga 3181 taccgtcgac ctcgacccgg gcggccgctt cgagcagaca tgataagata cattgatgag 3241 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat 3301 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc 3361 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac 3421 ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta 3481 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc 3541 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt 3601 tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc 3661 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc 3721 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc 3781 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc 3841 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 3901 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 3961 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 4021 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 4081 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 4141 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg 4201 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct 4261 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc 4321 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa 4381 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt 4441 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct 4501 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc 4561 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta 4621 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac 4681 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc 4741 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac 4801 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg 4861 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac 4921 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc 4981 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt 5041 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga 5101 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc 5161 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag 5221 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca 5281 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc 5341 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca 5401 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 5461 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 5521 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt 5581 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 5641 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 5701 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 5761 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 5821 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 5881 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 5941 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 6001 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 6061 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 6121 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca 6181 gtgagcgagg aagcggaaga //