LOCUS Exported 7233 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION vCre-Dependent ChR2(H134R)-eYFP with the vLox sites in the reverse orientation. ACCESSION . VERSION . KEYWORDS pAAV-Ef1a-vCreDIO hChR2(H134R)-eYFP 2.0 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7233) TITLE INTRSECT 2.0 JOURNAL Unpublished REFERENCE 2 (bases 1 to 7233) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7233 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 231..1409 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 462..1400 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1357..1377 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS complement(1546..2265) /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1591..1612) /label=EGFP-C /note="EGFP, forward primer" primer_bind 1940..1959 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2199..2220 /label=EGFP-N /note="EGFP, reverse primer" regulatory 2262..2271 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS complement(2281..3207) /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 3204..3213 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" misc_feature 3360..3948 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3413..3433) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3831..3842) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3980..4456 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4179..4198) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4496..4636 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4496..4625 /label=AAV2 ITR rep_origin 4711..5166 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4798..4817) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5008..5029 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5183..5202) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5302..5324 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5362..5380) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5448..5552 /gene="bla" /label=AmpR promoter CDS 5553..6413 /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(5771..5790) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6584..7172 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7073..7092 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga 181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg 241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg 301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt 361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg 421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt 481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct 541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga 601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct 661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt 721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc 781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg 841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac 901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc 961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag 1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct 1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg 1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga 1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc 1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg 1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa 1381 gtttttttct tccatttcag gtgtcgtgag gatccatcaa tttctgagaa ctgtcattct 1441 cggaaattga gtagctggag caatttgaaa agattgttgt attacctcta gctgcgctta 1501 tcaatttctg agaagtgtct ttctcggaaa ttgaggcgcg ccctattact tgtacagctc 1561 gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg 1621 cttctcgttg gggtctttgc tcagggcgga ctggtagctc aggtagtggt tgtcgggcag 1681 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct 1741 gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc 1801 ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt 1861 gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc 1921 gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc 1981 ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg 2041 gtagcgggcg aagcactgca ggccgtagcc gaaggtggtc acgagggtgg gccagggcac 2101 gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc 2161 ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag 2221 gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg cggccgctgg 2281 cacggctccg gcctcggctt cgtcttcgac gagagtctcg acctcgatct ccgttccgcc 2341 gatgttcagt ttggtggttt tgcggatatc tccgtgaatc aatatgtgct cgtggatcag 2401 gacgcgcagg tagtgtccca acaaccccca acaattttta ctcatcagat caataatcgt 2461 gtgacctacg gtggagccat agacgctcag gacgccaaaa ccttcgggcc ccaaaatgaa 2521 gagaattggg aacatacccc agctcacgaa aaacagccat gccatgccgg tcacgacctg 2581 gcggcaccga ccctttggca cagtatgata accctcgata tatgctttgg cggcgtgaaa 2641 aaatgtgttc gcgccatagc acaatccaag acaaaagaag atgactttaa catagccggt 2701 tgccatggcg ctggtagccc cccacacgat agtcccgatg tctgagacaa ggagtcccat 2761 ggttctcctg ctgtagtcgt tgctcaggcc ggtgaggttg ctcaggcgga taaggatgac 2821 aggacaagtg agcagccact ctgcatagcg cagccactgc acccggtgtc ctgtggcaag 2881 gtagagcata gagggattct taaactcaaa aaagaactcg agaatcacct taaccatttc 2941 aatggcgcac acatagatct cctcccagcc gcatgtagat ttccaggttt ggtaggcata 3001 gaacatcagc agcaaaatgc tgaatcctgc tgcaagccac tgcaggacat ttgacgcggt 3061 ctgagcgccg ttcgtgccgc gagattcaat ccatccggca cagtaacatt gatcctcagg 3121 gaccaggacg gacccgttca ccaccacagg attagtaacg aacaaaagtt cgcgtccgac 3181 ggcagacaaa gcgccgccat agtccatggt ggctagctca atttccgaga atgacagttc 3241 tcagaaattg ataagcagac aatcttgtaa tacatgcagc tagtcaagat tgctccagct 3301 atcaatttcc gagaaagaca cttctcagaa attgagaatt cgatatcaag cttatcgata 3361 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 3421 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 3481 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 3541 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 3601 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 3661 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 3721 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 3781 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 3841 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 3901 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag 3961 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc 4021 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat 4081 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa 4141 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag 4201 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct 4261 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt 4321 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc 4381 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc 4441 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa 4501 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg 4561 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg 4621 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt 4681 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg 4741 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc 4801 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 4861 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 4921 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 4981 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 5041 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt 5101 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta 5161 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc 5221 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt 5281 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac 5341 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga 5401 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta 5461 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat 5521 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc 5581 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga 5641 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca 5701 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt 5761 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg 5821 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc 5881 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata 5941 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt 6001 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag 6061 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca 6121 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg 6181 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg 6241 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag 6301 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg 6361 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag 6421 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga 6481 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt 6541 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc 6601 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 6661 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac 6721 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac 6781 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt 6841 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct 6901 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat 6961 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt 7021 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg 7081 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt 7141 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt 7201 tcctggcctt ttgctggcct tttgctcaca tgt //