LOCUS Exported 5895 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express the membrane targeted Y-GECO1 in neurons. ACCESSION . VERSION . KEYWORDS AAV-hSyn-Lyn-Y-GECO1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5895) AUTHORS Zhao Y, Abdelfattah AS, Zhao Y, Ruangkittisakul A, Ballanyi K, Campbell RE, Harrison DJ TITLE Microfluidic cell sorter-aided directed evolution of a protein-based calcium ion indicator with an inverted fluorescent response. JOURNAL Integr Biol (Camb). 2014 May 19. PUBMED 24840546 REFERENCE 2 (bases 1 to 5895) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5895 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 95..112 /label=L4440 /note="L4440 vector, forward primer" protein_bind 229..250 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 265..295 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 303..319 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 308..330 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 327..343 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 327..343 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" repeat_region 381..510 /label=AAV2 ITR promoter 592..1038 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 1055..1064 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2152..2166 /codon_start=1 /product="enterokinase recognition and cleavage site" /label=enterokinase site /translation="DDDDK" misc_feature 2405..2993 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2458..2478) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2876..2887) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2996..3012) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2997..3013) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 3042..3163 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3053..3072) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3107..3126 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 3325..3454 /label=AAV2 ITR primer_bind complement(3473..3490) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3473..3489) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3482..3504) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3631..4086 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3718..3737) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3928..3949 /label=F1ori-F /note="F1 origin, forward primer" promoter 4112..4216 /gene="bla" /label=AmpR promoter CDS 4217..5077 /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(4435..4454) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5248..5836 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5737..5756 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc 61 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga 121 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg 181 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta 241 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg 301 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccagatt 361 taattaaggc cttaattagg ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc 421 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg 481 gagtggccaa ctccatcact aggggttcct tgtagttaat gattaacccg ccatgctact 541 tatctacgta gccatgctct aggaagatct ctgcagaggg ccctgcgtat gagtgcaagt 601 gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg accccaccca 661 ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg 721 ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct 781 tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca 841 ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc 901 ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg 961 cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg 1021 tcgtgcctga gagcgcagtc gaattcgggg atccgccacc atgggatgta taaagagtaa 1081 aagaaaagat aacttaaatg atgacgaaga tgataaggat ctcgccacaa tggccgactc 1141 atcacgtcgt aagtggaata agacacacgc agtcagagct ataggccggc tgagctcact 1201 cgagggttat agtaccgaga aaattgtgcc tgttccaaag caagggatat taaaaggaaa 1261 ggtcagcatg tacctattgc tgaaggatgg cggtcgttat cattgccagt ttgatacggt 1321 atataaagct aaatcagtgc cctccaagat gccaggacgg cattttatac agcataagct 1381 ccttagggag gaccgctctg atgctaaaaa ccataaatgg caattaacag aacatgcaat 1441 cgcaggcatg gacgagctgt acaagggagg tacaggcggg agtatggtga gcaagggcga 1501 ggggcaatcg aagcatgggc taaaggagga gatgacggtg aagtaccaca tggaaggttg 1561 cgttaatggt cacaaatttg tcattactgg agagggcatt ggaaaccctt ttaagggtaa 1621 gcaaactgca aatttgtgtg tgatagaagg aggcccgctg ccgttctcgg aggacatttt 1681 aagtccgggt tttaaatatg gtgaccggat tttcacagag tacccacagg atattgtaga 1741 ttacttcaag aactcatgtc cagagggcta tacgtgggaa aggagcttct tctatgagga 1801 cggagcggtc tgtcgatgca acgcggacat aacagtctct gaaaaggaga actgcatcta 1861 tcacaaaagt atcttctatg gggtgaattt tcccgccgac ggccccgtaa tgaagaaaat 1921 gaccactaat tgggaagctg acacgcgtga ccaactgact gaagagcaga tcgcagaatt 1981 taaagaggct ttctccctat ttgacaagga cggggatggg acgatgacaa ccgaggagct 2041 ggggacggtg ctgcggtctc tggggcagaa cccctcagaa gcagagctgc gggacatgat 2101 caatgaagta gatgccgacg gtgacggcac aatcggcttc cctgagttcc tgacgatgat 2161 gacaagaaaa atgaatgaca cggacagtga agaggaaatt agagaagcgt tccgtgtgtt 2221 tgataaggac ggcaatggct acatcggcgc agcagagctt cgccacgtga tgacaaacct 2281 tggagagaag ttaacagatg aagagtttga tgaaatgatc agggtagcag actccgacgg 2341 ggatggtcag gtaaactacg aagagtttgt acaaatgatg acagcaaagt gaaagcttat 2401 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 2461 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 2521 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 2581 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 2641 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 2701 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 2761 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 2821 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 2881 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 2941 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata 3001 ccgtcgacct cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt 3061 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc 3121 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat 3181 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct 3241 ctacaaatgt ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc 3301 atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc 3361 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg 3421 cccgggcggc ctcagtgagc gagcgagcgc gcagccttaa ttaacctaat tcactggccg 3481 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag 3541 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc 3601 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg 3661 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 3721 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 3781 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 3841 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 3901 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 3961 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt 4021 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta 4081 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 4141 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 4201 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 4261 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 4321 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 4381 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 4441 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 4501 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 4561 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 4621 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 4681 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 4741 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga 4801 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 4861 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 4921 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 4981 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 5041 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 5101 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 5161 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 5221 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 5281 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 5341 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct 5401 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 5461 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 5521 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 5581 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 5641 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 5701 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 5761 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 5821 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 5881 gccttttgct cacat //