LOCUS Exported 7457 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses Flpo recombinase-dependent GtACR2-Kv2.1C-TlcnC, which targets GtACR2 to the somatodendritic compartment. ACCESSION . VERSION . KEYWORDS pAAV-EF1-alpha-FRT-FLEX-GtACR2-EYFP-Kv2.1C-TlcnC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7457) 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 7457) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7457 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 122..251 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 352..1530 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 583..1521 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1478..1498 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 1621..1654 /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(1929..2642) /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(1971..1992) /label=EGFP-C /note="EGFP, forward primer" primer_bind 2320..2339 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2579..2600 /label=EGFP-N /note="EGFP, reverse primer" protein_bind complement(3645..3678) /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 3705..4293 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3758..3778) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4176..4187) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4325..4801 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4524..4543) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4841..4981 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4841..4970 /label=AAV2 ITR rep_origin 5056..5511 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5143..5162) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5353..5374 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5528..5547) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5647..5669 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5707..5725) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5793..5897 /gene="bla" /label=AmpR promoter CDS 5898..6758 /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(6116..6135) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6929..60 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7418..7437 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 61 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 121 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt 181 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac 241 taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt ttaaacagag 301 aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt 361 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc 421 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg 481 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc 541 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt 601 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc 661 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg 721 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc 781 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag 841 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt 901 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc 961 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca 1021 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg 1081 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga 1141 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc 1201 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc 1261 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg 1321 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc 1381 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg 1441 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa 1501 agtttttttc ttccatttca ggtgtcgtga ggtaccgaag ttcctattct atcagaagta 1561 taggaacttc gcagaatggt agctggattg tagctgctat tagcaatatg aaacctctta 1621 gaagttccta ttctctagaa agtataggaa cttcgaattc ttacttgtac aggcgcgcct 1681 caggaagatg tcagctggat agcgaaaacc tcgccatctg caggtgactc tgcaaacctt 1741 gtagcttcgg ggaagtcggt ggcgcaggaa atgaagctat cgatactaga catgcttctc 1801 atgtcaatca cgccctcggt cctagctggc agaggagcca ctggagatgg catggagctc 1861 atttccagtt cctcaggggg tttactctga ggggccatct ccttagtgtt caggatgggc 1921 tgagactgct tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc 1981 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctggtagctc 2041 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag 2101 tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg 2161 atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc 2221 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg 2281 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc 2341 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg 2401 tgctgcttca tgtggtcggg gtagcgggcg aagcactgca ggccgtagcc gaaggtggtc 2461 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc 2521 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg 2581 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc 2641 acggcggccg ccctgccgaa cattctgatg ttgacctcgt cgttgggggg ggcattcttc 2701 tcgtggttca gtccgaaggg gtcgtcggcc tcaggcttct cctggccagg gaggcaagcc 2761 gggtcccact ttccctcgag cactccccag gcggtggacc acaggatcac gccgtaggtg 2821 ttcttgcaca ggccgtcggc gagcagatac aggatggcgc tcacgttctc gtggatcacg 2881 cacacgccgg tgggggaaat cagccacacg atggggaaca tggtccaggc gatgaagaag 2941 atcagcctca gcatcttgat cctcaccacc accttctggg ccagaggggt gcccacggac 3001 tcgaagccgt acagtcccac ctggaagatg gtgaacacca cgctgtattc gaagaccagg 3061 caggtagcgc cgaagaagaa gaacagccac ttcatgtact cggcggggga gatggtagcg 3121 gtaatgccca tcaccaccct gataatgctg gcggcctggg cgatggggtt cagagggatg 3181 ctcttgtact tcaccagagc catgttggag atctggccca gcataatggg gcatgtgcac 3241 agccagctgg ccatcctgga ccagggaaag aacctgccgt ccctcatctg cagtgtgcca 3301 ttgccggtgc tggccagggc gtatgtgatg gactccacga agggcaggta cacgctctcc 3361 cagggcacct tgggggccct gctgatcatg aagatggtct ggcatccgct caccacggcc 3421 cacaccagct gcagcaggct cacgaaggtg ctgtcgatcc tgctcatgtt gtagcactct 3481 gtgtgggtgc tggcccactc gccgtacacg acctgggagg ccatggatcc cggctagcta 3541 cgtaggcgcg ccttaattaa gaagttccta tacttctgat agaataggaa cttcttgcct 3601 taacccagaa attatcactg ttattcttta gaatggtgca aagagaagtt cctatacttt 3661 ctagagaata ggaacttcca gaattcgata tcaagcttat cgataatcaa cctctggatt 3721 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 3781 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 3841 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 3901 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 3961 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 4021 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 4081 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct 4141 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 4201 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 4261 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat 4321 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac 4381 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt 4441 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa 4501 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc 4561 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga 4621 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag 4681 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc 4741 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct 4801 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag 4861 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc 4921 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag 4981 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt 5041 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 5101 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc 5161 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt 5221 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg 5281 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 5341 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct 5401 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga 5461 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca 5521 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac 5581 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga 5641 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac 5701 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt 5761 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 5821 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 5881 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 5941 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 6001 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 6061 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat 6121 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact 6181 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 6241 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 6301 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 6361 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 6421 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg 6481 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 6541 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 6601 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 6661 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 6721 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat 6781 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 6841 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 6901 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 6961 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 7021 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc 7081 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 7141 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 7201 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 7261 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 7321 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 7381 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 7441 gtcctgtcgg gtttcgc //