LOCUS Exported 7413 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION expresses Flpo recombinase-dependent GtACR2-Kv2.1C, which targets GtACR2 to the somatodendritic compartment. ACCESSION . VERSION . KEYWORDS pAAV-EF1-alpha-FRT-FLEX-GtACR2-EYFP-Kv2.1C SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7413) AUTHORS Messier JE, Chen H, Cai ZL, Xue M TITLE Targeting light-gated chloride channels to neuronal somatodendritic domain reduces their excitatory effect in the axon. JOURNAL Elife. 2018 Aug 9;7. pii: 38506. doi: 10.7554/eLife.38506. PUBMED 30091701 REFERENCE 2 (bases 1 to 7413) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7413 /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" protein_bind 1510..1543 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" CDS complement(1764..2477) /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(1806..1827) /label=EGFP-C /note="EGFP, forward primer" primer_bind 2155..2174 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2414..2435 /label=EGFP-N /note="EGFP, reverse primer" protein_bind complement(3480..3513) /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" misc_feature 3540..4128 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3593..3613) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4011..4022) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4160..4636 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4359..4378) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4676..4816 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4676..4805 /label=AAV2 ITR rep_origin 4891..5346 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4978..4997) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5188..5209 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5363..5382) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5482..5504 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5542..5560) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5628..5732 /gene="bla" /label=AmpR promoter CDS 5733..6593 /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(5951..5970) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6764..7352 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7253..7272 /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 gtaccgaagt tcctattcta 1441 tcagaagtat aggaacttcg cagaatggta gctggattgt agctgctatt agcaatatga 1501 aacctcttag aagttcctat tctctagaaa gtataggaac ttcgaattct tacttgtacc 1561 gcgccttaaa accttgtagc ttcggggaag tcggtggcgc aggaaatgaa gctatcgata 1621 ctagacatgc ttctcatgtc aatcacgccc tcggtcctag ctggcagagg agccactgga 1681 gatggcatgg agctcatttc cagttcctca gggggtttac tctgaggggc catctcctta 1741 gtgttcagga tgggctgaga ctgcttgtac agctcgtcca tgccgagagt gatcccggcg 1801 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg 1861 gcggactggt agctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg 1921 gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc 1981 ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg 2041 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc 2101 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg 2161 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg 2221 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca ctgcaggccg 2281 tagccgaagg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg 2341 aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac 2401 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc 2461 tcctcgccct tgctcacggc ggccgccctg ccgaacattc tgatgttgac ctcgtcgttg 2521 gggggggcat tcttctcgtg gttcagtccg aaggggtcgt cggcctcagg cttctcctgg 2581 ccagggaggc aagccgggtc ccactttccc tcgagcactc cccaggcggt ggaccacagg 2641 atcacgccgt aggtgttctt gcacaggccg tcggcgagca gatacaggat ggcgctcacg 2701 ttctcgtgga tcacgcacac gccggtgggg gaaatcagcc acacgatggg gaacatggtc 2761 caggcgatga agaagatcag cctcagcatc ttgatcctca ccaccacctt ctgggccaga 2821 ggggtgccca cggactcgaa gccgtacagt cccacctgga agatggtgaa caccacgctg 2881 tattcgaaga ccaggcaggt agcgccgaag aagaagaaca gccacttcat gtactcggcg 2941 ggggagatgg tagcggtaat gcccatcacc accctgataa tgctggcggc ctgggcgatg 3001 gggttcagag ggatgctctt gtacttcacc agagccatgt tggagatctg gcccagcata 3061 atggggcatg tgcacagcca gctggccatc ctggaccagg gaaagaacct gccgtccctc 3121 atctgcagtg tgccattgcc ggtgctggcc agggcgtatg tgatggactc cacgaagggc 3181 aggtacacgc tctcccaggg caccttgggg gccctgctga tcatgaagat ggtctggcat 3241 ccgctcacca cggcccacac cagctgcagc aggctcacga aggtgctgtc gatcctgctc 3301 atgttgtagc actctgtgtg ggtgctggcc cactcgccgt acacgacctg ggaggccatg 3361 gatcccggct agctacgtag gcgcgcctta attaagaagt tcctatactt ctgatagaat 3421 aggaacttct tgccttaacc cagaaattat cactgttatt ctttagaatg gtgcaaagag 3481 aagttcctat actttctaga gaataggaac ttccagaatt cgatatcaag cttatcgata 3541 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 3601 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 3661 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 3721 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 3781 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 3841 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 3901 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 3961 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 4021 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 4081 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag 4141 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc 4201 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat 4261 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa 4321 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag 4381 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct 4441 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt 4501 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc 4561 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc 4621 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa 4681 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg 4741 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg 4801 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt 4861 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg 4921 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 4981 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 5041 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 5101 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 5161 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 5221 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt 5281 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta 5341 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc 5401 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt 5461 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac 5521 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga 5581 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta 5641 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat 5701 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc 5761 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga 5821 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca 5881 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt 5941 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg 6001 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc 6061 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata 6121 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt 6181 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag 6241 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca 6301 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg 6361 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg 6421 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag 6481 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg 6541 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag 6601 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga 6661 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt 6721 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc 6781 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 6841 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac 6901 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac 6961 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt 7021 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct 7081 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat 7141 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt 7201 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg 7261 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt 7321 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt 7381 tcctggcctt ttgctggcct tttgctcaca tgt //