LOCUS Exported 6394 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV-mediated expression of Chronos-GFP under the Syn promoter, in floxed/reversed (Cre-dependent) manner. Using bGHpA signal.. ACCESSION . VERSION . KEYWORDS pAAV-Syn-FLEX-rc[Chronos-GFP] SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6394) AUTHORS Klapoetke NC, Murata Y, Kim SS, Pulver SR, Birdsey-Benson A, Cho YK, Morimoto TK, Chuong AS, Carpenter EJ, Tian Z, Wang J, Xie Y, Yan Z, Zhang Y, Chow BY, Surek B, Melkonian M, Jayaraman V, Constantine-Paton M, Wong GK, Boyden ES TITLE Independent optical excitation of distinct neural populations. JOURNAL Nat Methods. 2014 Mar;11(3):338-46. doi: 10.1038/nmeth.2836. Epub 2014 Feb 9. PUBMED 24509633 REFERENCE 2 (bases 1 to 6394) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6394 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(11..28) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(182..770) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(262..281) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(941..1801) /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 1564..1583 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1802..1906) /gene="bla" /label=AmpR promoter rep_origin complement(1932..2387) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2069..2090) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2281..2300 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 2459..2588 /label=AAV2 ITR promoter 2691..3138 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" protein_bind complement(3154..3187) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(3250..3283) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(3300..4013) /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(3342..3363) /label=EGFP-C /note="EGFP, forward primer" primer_bind 3691..3710 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3950..3971 /label=EGFP-N /note="EGFP, reverse primer" regulatory 5003..5012 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind 5034..5067 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 5138..5171 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 5204..5792 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5257..5277) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5675..5686) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 5833..6040 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 6210..6339 /label=AAV2 ITR ORIGIN 1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta 1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga 2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa 2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga 2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc 2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga 2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg 2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt 2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct 2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag 2641 gaagatcgta ggttaattaa gtgtctagac tgcagagggc cctgcgtatg agtgcaagtg 2701 ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca 2761 ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg 2821 ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct 2881 tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca 2941 ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc 3001 ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg 3061 cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg 3121 tcgtgcctga gagcgcagtc gagaagaatt cccataactt cgtataaagt atcctatacg 3181 aagttatatc aaaataggaa gaccaatgct tcaccatcga cccgaattgc caagcatcac 3241 catcgaccca taacttcgta taatgtatgc tatacgaagt tatactagct agctcattac 3301 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc 3361 atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg 3421 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg 3481 agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc 3541 ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc 3601 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg 3661 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag 3721 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc 3781 atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg 3841 ggccagggca cgggcagctt gccggtggtg caaatgaact tcagggtcag cttgccgtag 3901 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc 3961 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct cactgctact 4021 accggtgccg ccactcctcc ctcctcctcc tcgtcgacga aggtctcgat ctccatgttc 4081 tcgccggcca cgttgatggt ggttgtcttc cggatgtcgc cgtggatcag gatgtgctcg 4141 tggatcttca ctctcaggtg gtggcccagg aagccccaag tgttcttgct gatcagatcc 4201 aggatcaggt ggccgattcc gctggtgtaa ggtgtgatca ggcccagtcc ctcgtgtccg 4261 gcgatgaaca tcacggggaa catcagccaa gagcagaaga agacgtaggc catgatcttg 4321 caaatcttcc ggcagacgcc tttgggcagg gtgtggtagc tctcgatata caccttggcg 4381 atctggaaga agcaagtcac gccgtagaac aggccgatca tgaagaacag gatcttcagg 4441 gggccctttg taaaggcggc tgtgatcccc cacacgatgt tgccgatgtc ggtcaccagg 4501 atggtcatgg tccgcttgct gtactcttcg tgcagtccgg tcaggttgct caggtggatc 4561 aggatcacgg ggcaagtcag gagccacatg ctgtaccgca gccaaatcac ggctcctccg 4621 ttggtctggt acactgtggc tgggctgtcg acctcgtgga acagctcgat gaagcacttg 4681 acgagctcga tcacgcagac gtagacctcc tcccagccgg ttgtggcctt gaagctgtgc 4741 caaccgtaga agatgagctg cacgatagcc acgatcacgg tgaaccactg gagtcccact 4801 gcgatcttgt gctgcagctc ggttccgtgg ttgatgtggg ggaagcaacc gtgatcggct 4861 ccggctgttg tggcgttaga ggtttctccg tgagcgtctg ctgctggtgt caccactgct 4921 gcggcgctca gcaggcctct aattgtggct tcggcgctag gcacggctga gataaaggcg 4981 tgggtcattg tggcggctgt ttccatggtg gcggcgcggc ctaggtacct agtataactt 5041 cgtataggat actttatacg aagttatcat tgggattctt cctattttga tccaagcatc 5101 accatcgacc ctctagtcca gatctcacca tcgacccata acttcgtata gcatacatta 5161 tacgaagtta tgtccctcga agaggttcgc gaagcttatc gataatcaac ctctggatta 5221 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg 5281 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc 5341 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca 5401 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac 5461 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact 5521 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc 5581 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg 5641 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc 5701 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac 5761 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacccg ggcggccgct 5821 cataaagctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc 5881 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg 5941 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca 6001 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtccgg actgtacaat 6061 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa 6121 aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa 6181 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc 6241 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg 6301 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcattaatg aatcggccaa 6361 cgcgcgggga gaggcggttt gcgtattggg cgct //