LOCUS Exported 7249 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION sCre-Dependent ChR2-EYFP. ACCESSION . VERSION . KEYWORDS pAAV-Ef1a-sCreDIO hChR2(H134R)-EYFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7249) AUTHORS Fenno LE, Mattis J, Ramakrishnan C, Hyun M, Lee SY, He M, Tucciarone J, Selimbeyoglu A, Berndt A, Grosenick L, Zalocusky KA, Bernstein H, Swanson H, Perry C, Diester I, Boyce FM, Bass CE, Neve R, Huang ZJ, Deisseroth K TITLE Targeting cells with single vectors using multiple-feature Boolean logic. JOURNAL Nat Methods. 2014 Jul;11(7):763-72. doi: 10.1038/nmeth.2996. Epub 2014 Jun 8. PUBMED 24908100 REFERENCE 2 (bases 1 to 7249) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7249 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 12..141 /label=AAV2 ITR promoter 242..1419 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 473..1410 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1367..1387 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS complement(1562..2281) /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1607..1628) /label=EGFP-C /note="EGFP, forward primer" primer_bind 1956..1975 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2215..2236 /label=EGFP-N /note="EGFP, reverse primer" regulatory 2278..2287 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS complement(2297..3223) /codon_start=1 /product="H134R mutant of an N-terminal fragment of channelrhodopsin-2 from the alga Chlamydomonas reinhardtii (Nagel et al., 2005)" /label=ChR2(H134R) /note="blue light-activated cation channel" /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI RKTTKLNIGGTEIEVETLVEDEAEAGAV" regulatory 3220..3229 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" misc_feature 3376..3964 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3429..3449) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3847..3858) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3996..4472 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4195..4214) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4512..4652 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4512..4641 /label=AAV2 ITR rep_origin 4727..5182 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4814..4833) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5024..5045 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5199..5218) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5318..5340 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5378..5396) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5464..5568 /gene="bla" /label=AmpR promoter CDS 5569..6429 /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(5787..5806) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6600..7188 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7089..7108 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt 181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat 241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg 301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag 361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc 421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc 481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat 541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc cactcgtgtc 1441 cgataactgt aattatcgga catgataatt gttattggtg caaagaatca cttccttaag 1501 tctttatact gcccagctcg tgtccgataa gtgtatttat cggacatgat ggcgcgccct 1561 attacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca 1621 ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag ggcggactgg tagctcaggt 1681 agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt 1741 cggcgagctg cacgctgccg tcctcgatgt tgtggcggat cttgaagttc accttgatgc 1801 cgttcttctg cttgtcggcc atgatataga cgttgtggct gttgtagttg tactccagct 1861 tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca 1921 ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga 1981 agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct 2041 gcttcatgtg gtcggggtag cgggcgaagc actgcaggcc gtagccgaag gtggtcacga 2101 gggtgggcca gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc 2161 cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc 2221 cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca 2281 tggtggcggc cgctggcacg gctccggcct cggcttcgtc ttcgacgaga gtctcgacct 2341 cgatctccgt tccgccgatg ttcagtttgg tggttttgcg gatatctccg tgaatcaata 2401 tgtgctcgtg gatcaggacg cgcaggtagt gtcccaacaa cccccaacaa tttttactca 2461 tcagatcaat aatcgtgtga cctacggtgg agccatagac gctcaggacg ccaaaacctt 2521 cgggccccaa aatgaagaga attgggaaca taccccagct cacgaaaaac agccatgcca 2581 tgccggtcac gacctggcgg caccgaccct ttggcacagt atgataaccc tcgatatatg 2641 ctttggcggc gtgaaaaaat gtgttcgcgc catagcacaa tccaagacaa aagaagatga 2701 ctttaacata gccggttgcc atggcgctgg tagcccccca cacgatagtc ccgatgtctg 2761 agacaaggag tcccatggtt ctcctgctgt agtcgttgct caggccggtg aggttgctca 2821 ggcggataag gatgacagga caagtgagca gccactctgc atagcgcagc cactgcaccc 2881 ggtgtcctgt ggcaaggtag agcatagagg gattcttaaa ctcaaaaaag aactcgagaa 2941 tcaccttaac catttcaatg gcgcacacat agatctcctc ccagccgcat gtagatttcc 3001 aggtttggta ggcatagaac atcagcagca aaatgctgaa tcctgctgca agccactgca 3061 ggacatttga cgcggtctga gcgccgttcg tgccgcgaga ttcaatccat ccggcacagt 3121 aacattgatc ctcagggacc aggacggacc cgttcaccac cacaggatta gtaacgaaca 3181 aaagttcgcg tccgacggca gacaaagcgc cgccatagtc catggtggct agcatcatgt 3241 ccgataaata cacttatcgg acacgagctg ggcagtataa agacttaagg aagtgattct 3301 ttgcaccaat aacaattatc atgtccgata attacagtta tcggacacga ggaattcgat 3361 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt 3421 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 3481 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 3541 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 3601 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 3661 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 3721 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc 3781 tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac 3841 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg 3901 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc 3961 ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc 4021 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa 4081 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg 4141 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt 4201 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg 4261 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca 4321 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg 4381 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac 4441 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac 4501 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc 4561 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc 4621 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac 4681 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc 4741 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 4801 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 4861 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 4921 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag 4981 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 5041 actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg 5101 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac 5161 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca 5221 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg 5281 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg 5341 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta 5401 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat 5461 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg 5521 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa 5581 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac 5641 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac 5701 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt 5761 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc 5821 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca 5881 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc 5941 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag 6001 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa 6061 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg 6121 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa 6181 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg 6241 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt 6301 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt 6361 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag 6421 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat 6481 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct 6541 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 6601 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 6661 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 6721 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc 6781 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 6841 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 6901 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 6961 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg 7021 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 7081 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 7141 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 7201 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt //