LOCUS Exported 6201 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 6201) 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 (iAChSnFR) JOURNAL Unpublished REFERENCE 2 (bases 1 to 6201) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6201 /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)" 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" rep_origin 3814..4269 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3901..3920) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4111..4132 /label=F1ori-F /note="F1 origin, forward primer" promoter 4295..4399 /gene="bla" /label=AmpR promoter CDS 4400..5260 /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(4618..4637) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5431..6019 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5920..5939 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6173..6190 /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 cgccatcatg 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 gcgagcsrsc gyryagysrg tcgtawtaca attcactggc 3661 cgycswytka gsccggssna ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct 3721 cagtgagcga gcgagcgcgc agctggcgta atagcgaaga ggcccgcacc gatcgccctt 3781 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg 3841 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 3901 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 3961 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 4021 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc 4081 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac 4141 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt 4201 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc 4261 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 4321 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 4381 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt 4441 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc 4501 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat 4561 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct 4621 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca 4681 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg 4741 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa 4801 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg 4861 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga 4921 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg 4981 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt 5041 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg 5101 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc 5161 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca 5221 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc 5281 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 5341 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 5401 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 5461 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 5521 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct 5581 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 5641 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 5701 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 5761 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 5821 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 5881 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 5941 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 6001 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 6061 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 6121 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 6181 agtgagcgag gaagcggaag a //