LOCUS Exported 7931 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pAAV-GFAP-hChR2(H134R)-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7931) TITLE hChR2 plasmids REFERENCE 2 (bases 1 to 7931) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7931 /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 regulatory 2381..2390 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2387..3313 /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" CDS 3326..4033 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE GRHSTGGMDELYK" primer_bind complement(3468..3486) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3736..3755 /label=mCherry-F /note="mCherry, forward primer" misc_feature 4058..4646 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4111..4131) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4529..4540) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4678..5154 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4877..4896) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 5194..5334 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 5194..5323 /label=AAV2 ITR rep_origin 5409..5864 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5496..5515) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5706..5727 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5881..5900) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6000..6022 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(6060..6078) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6146..6250 /gene="bla" /label=AmpR promoter CDS 6251..7111 /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(6469..6488) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7282..7870 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7771..7790 /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 cgcgtcccac ctccctctct gtgctgggac 181 tcacagaggg agacctcagg aggcagtctg tccatcacat gtccaaatgc agagcatacc 241 ctgggctggg cgcagtggcg cacaactgta attccagcac tttgggaggc tgatgtggaa 301 ggatcacttg agcccagaag ttctagacca gcctgggcaa catggcaaga ccctatctct 361 acaaaaaaag ttaaaaaatc agccacgtgt ggtgacacac acctgtagtc ccagctattc 421 aggaggctga ggtgagggga tcacttaagg ctgggaggtt gaggctgcag tgagtcgtgg 481 ttgcgccact gcactccagc ctgggcaaca gtgagaccct gtctcaaaag acaaaaaaaa 541 aaaaaaaaaa aaaaagaaca tatcctggtg tggagtaggg gacgctgctc tgacagaggc 601 tcgggggcct gagctggctc tgtgagctgg ggaggaggca gacagccagg ccttgtctgc 661 aagcagacct ggcagcattg ggctggccgc cccccagggc ctcctcttca tgcccagtga 721 atgactcacc ttggcacaga cacaatgttc ggggtgggca cagtgcctgc ttcccgccgc 781 accccagccc ccctcaaatg ccttccgaga agcccattga gcagggggct tgcattgcac 841 cccagcctga cagcctggca tcttgggata aaagcagcac agccccctag gggctgccct 901 tgctgtgtgg cgccaccggc ggtggagaac aaggctctat tcagcctgtg cccaggaaag 961 gggatcaggg gatgcccagg catggacagt gggtggcagg gggggagagg agggctgtct 1021 gcttcccaga agtccaagga cacaaatggg tgaggggact gggcagggtt ctgaccctgt 1081 gggaccagag tggagggcgt agatggacct gaagtctcca gggacaacag ggcccaggtc 1141 tcaggctcct agttgggccc agtggctcca gcgtttccaa acccatccat ccccagaggt 1201 tcttcccatc tctccaggct gatgtgtggg aactcgagga aataaatctc cagtgggaga 1261 cggaggggtg gccagggaaa cggggcgctg caggaataaa gacgagccag cacagccagc 1321 tcatgtgtaa cggctttgtg gagctgtcaa ggcctggtct ctgggagaga ggcacaggga 1381 ggccagacaa ggaaggggtg acctggaggg acagatccag gggctaaagt cctgataagg 1441 caagagagtg ccggccccct cttgccctat caggacctcc actgccacat agaggccatg 1501 attgaccctt agacaaaggg ctggtgtcca atcccagccc ccagccccag aactccaggg 1561 aatgaatggg cagagagcag gaatgtggga catctgtgtt caagggaagg actccaggag 1621 tctgctggga atgaggccta gtaggaaatg aggtggccct tgagggtaca gaacaggttc 1681 attcttcgcc aaattcccag caccttgcag gcacttacag ctgagtgaga taatgcctgg 1741 gttatgaaat caaaaagttg gaaagcaggt cagaggtcat ctggtacagc ccttccttcc 1801 cttttttttt tttttttttt gtgagacaag gtctctctct gttgcccagg ctggagtggc 1861 gcaaacacag ctcactgcag cctcaaccta ctgggctcaa gcaatcctcc agcctcagcc 1921 tcccaaagtg ctgggattac aagcatgagc caccccactc agccctttcc ttccttttta 1981 attgatgcat aataattgta agtattcatc atggtccaac caaccctttc ttgacccacc 2041 ttcctagaga gagggtcctc ttgcttcagc ggtcagggcc ccagacccat ggtctggctc 2101 caggtaccac ctgcctcatg caggagttgg cgtgcccagg aagctctgcc tctgggcaca 2161 gtgacctcag tggggtgagg ggagctctcc ccatagctgg gctgcggccc aaccccaccc 2221 cctcaggcta tgccaggggg tgttgccagg ggcacccggg catcgccagt ctagcccact 2281 ccttcataaa gccctcgcat cccaggagcg agcagagcca gagcaggttg gagaggagac 2341 gcatcacctc cgctgctcgc cgggatcctc tagagtcgac gccaccatgg actatggcgg 2401 cgctttgtct gccgtcggac gcgaactttt gttcgttact aatcctgtgg tggtgaacgg 2461 gtccgtcctg gtccctgagg atcaatgtta ctgtgccgga tggattgaat ctcgcggcac 2521 gaacggcgct cagaccgcgt caaatgtcct gcagtggctt gcagcaggat tcagcatttt 2581 gctgctgatg ttctatgcct accaaacctg gaaatctaca tgcggctggg aggagatcta 2641 tgtgtgcgcc attgaaatgg ttaaggtgat tctcgagttc ttttttgagt ttaagaatcc 2701 ctctatgctc taccttgcca caggacaccg ggtgcagtgg ctgcgctatg cagagtggct 2761 gctcacttgt cctgtcatcc ttatccgcct gagcaacctc accggcctga gcaacgacta 2821 cagcaggaga accatgggac tccttgtctc agacatcggg actatcgtgt ggggggctac 2881 cagcgccatg gcaaccggct atgttaaagt catcttcttt tgtcttggat tgtgctatgg 2941 cgcgaacaca ttttttcacg ccgccaaagc atatatcgag ggttatcata ctgtgccaaa 3001 gggtcggtgc cgccaggtcg tgaccggcat ggcatggctg tttttcgtga gctggggtat 3061 gttcccaatt ctcttcattt tggggcccga aggttttggc gtcctgagcg tctatggctc 3121 caccgtaggt cacacgatta ttgatctgat gagtaaaaat tgttgggggt tgttgggaca 3181 ctacctgcgc gtcctgatcc acgagcacat attgattcac ggagatatcc gcaaaaccac 3241 caaactgaac atcggcggaa cggagatcga ggtcgagact ctcgtcgaag acgaagccga 3301 ggccggagcc gtgccagcgg ccgccgtgag caagggcgag gaggataaca tggccatcat 3361 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga 3421 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt 3481 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg 3541 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc 3601 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac 3661 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa 3721 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga 3781 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa 3841 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca 3901 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta 3961 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga 4021 gctgtacaag taagaattcg atatcaagct tatcgataat caacctctgg attacaaaat 4081 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc 4141 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt 4201 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg 4261 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg 4321 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc 4381 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt 4441 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct 4501 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg 4561 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg 4621 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg 4681 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg 4741 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc 4801 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc 4861 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc 4921 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg 4981 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt 5041 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg 5101 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt 5161 ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca 5221 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc 5281 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc 5341 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca 5401 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag 5461 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt 5521 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt 5581 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg 5641 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt 5701 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt 5761 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca 5821 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag 5881 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga 5941 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc 6001 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg 6061 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc 6121 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca 6181 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa 6241 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt 6301 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca 6361 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag 6421 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc 6481 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca 6541 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt 6601 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct 6661 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt 6721 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga 6781 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact 6841 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc 6901 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga 6961 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt 7021 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga 7081 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact 7141 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga 7201 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 7261 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 7321 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 7381 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta 7441 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 7501 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 7561 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 7621 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 7681 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 7741 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 7801 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 7861 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 7921 tgctcacatg t //