LOCUS Exported 6970 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cre-on/Flp-on ChR2-EYFP under the short Ef1a promoter. ACCESSION . VERSION . KEYWORDS pAAV-nEF Con/Fon hChR2(H134R)-EYFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6970) 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 6970) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6970 /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 188..399 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 412..680 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" CDS complement(837..1088) /codon_start=1 /product="C-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /label=VC155 /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK" primer_bind complement(879..900) /label=EGFP-C /note="EGFP, forward primer" primer_bind 1228..1247 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" protein_bind 1469..1502 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 1553..1586 /label=loxN /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GTATACCT). loxN sites are compatible with each other, but incompatible with loxP or lox2272 sites." primer_bind complement(2252..2273) /label=EGFP-N /note="EGFP, reverse primer" protein_bind complement(2386..2419) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(2470..2503) /label=loxN /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GTATACCT). loxN sites are compatible with each other, but incompatible with loxP or lox2272 sites." regulatory 2939..2948 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" misc_feature 3097..3685 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3150..3170) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3568..3579) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3717..4193 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3916..3935) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4233..4373 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4233..4362 /label=AAV2 ITR rep_origin 4448..4903 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4535..4554) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4745..4766 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4920..4939) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5039..5061 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5099..5117) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5185..5289 /gene="bla" /label=AmpR promoter CDS 5290..6150 /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(5508..5527) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6321..6909 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6810..6829 /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 cgcgtaagga tctgcgatcg ctccggtgcc 181 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 241 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 301 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 361 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 421 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 481 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 541 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 601 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 661 gttctgcgcc gttacagatc caagctgtga ccggcgccta cggatccgaa gttcctattc 721 ttcaaatagt ataggaactt cgcagaatgg tagctggatt gtagctgcta ttagcaatat 781 gaaacctctt agaagttcct attcttcaaa aggtatagga acttcggcgc gccttacttg 841 tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg 901 tgatcgcgct tctcgttggg gtctttgctc agggcggact ggtagctcag gtagtggttg 961 tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc 1021 tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat gccgttcttc 1081 tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc 1141 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg 1201 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg 1261 gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg 1321 tggtcggggt agcgggcgaa gcactgcagg ccgtagccga aggtggtcac gagggtgggc 1381 cagggcacgg gcagctgtgg gacgacatga cttaaccagt taaccatgtt ctctgaacat 1441 gagaagggct ggccactctc cagtcgacat aacttcgtat aggatacttt atacgaagtt 1501 atgcagaatg gtagctggat tgtagctgct attagcaata tgaaacctct taataacttc 1561 gtatagtata ccttatacga agttatacta gttgtgcccc cccttttttt tataaattgt 1621 attaatgtta tatacatatc tcctgtatgt gacccatgtg cttatgactc tatttctcat 1681 gtgtttaggc ctgagcaacg actacagcag gagaaccatg ggactccttg tctcagacat 1741 cgggactatc gtgtgggggg ctaccagcgc catggcaacc ggctatgtta aagtcatctt 1801 cttttgtctt ggattgtgct atggcgcgaa cacatttttt cacgccgcca aagcatatat 1861 cgagggttat catactgtgc caaagggtcg gtgccgccag gtcgtgaccg gcatggcatg 1921 gctgtttttc gtgagctggg gtatgttccc aattctcttc attttggggc ccgaaggttt 1981 tggcgtcctg agcgtctatg gctccaccgt aggtcacacg attattgatc tgatgagtaa 2041 aaattgttgg gggttgttgg gacactacct gcgcgtcctg atccacgagc acatattgat 2101 tcacggagat atccgcaaaa ccaccaaact gaacatcggc ggaacggaga tcgaggtcga 2161 gactctcgtc gaagacgaag ccgaggccgg agccgtgcca gcggccgccg tgagcaaggg 2221 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg 2281 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct 2341 gaagttcatc tgcaccaccg gcaaggtgag tacaggaggg gtaccataac ttcgtataaa 2401 gtatcctata cgaagttatt tgccttaacc cagaaattat cactgttatt ctttagaatg 2461 gtgcaaagaa taacttcgta taaggtatac tatacgaagt tatgctagca accccgacac 2521 ttaccggtga ggttgctcag gcggataagg atgacaggac aagtgagcag ccactctgca 2581 tagcgcagcc actgcacccg gtgtcctgtg gcaaggtaga gcatagaggg attcttaaac 2641 tcaaaaaaga actcgagaat caccttaacc atttcaatgg cgcacacata gatctcctcc 2701 cagccgcatg tagatttcca ggtttggtag gcatagaaca tcagcagcaa aatgctgaat 2761 cctgctgcaa gccactgcag gacatttgac gcggtctgag cgccgttcgt gccgcgagat 2821 tcaatccatc cggcacagta acattgatcc tcagggacca ggacggaccc gttcaccacc 2881 acaggattag taacgaacaa aagttcgcgt ccgacggcag acaaagcgcc gccatagtcc 2941 atggtggctc tagagaagtt cctatactat ttgaagaata ggaacttctt gccttaaccc 3001 agaaattatc actgttattc tttagaatgg tgcaaagaga agttcctata ccttttgaag 3061 aataggaact tcgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt 3121 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 3181 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 3241 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 3301 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 3361 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 3421 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 3481 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg 3541 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 3601 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 3661 atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt 3721 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc 3781 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct 3841 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct 3901 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca 3961 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg 4021 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt 4081 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc 4141 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt 4201 tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac 4261 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc 4321 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct 4381 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa 4441 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc 4501 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt 4561 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc 4621 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt 4681 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt 4741 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt 4801 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa 4861 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt 4921 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac 4981 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc 5041 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc 5101 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca 5161 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat 5221 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa 5281 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt 5341 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag 5401 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt 5461 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg 5521 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag 5581 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta 5641 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg 5701 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta 5761 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac 5821 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt 5881 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca 5941 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag 6001 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta 6061 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag 6121 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt 6181 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat 6241 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta 6301 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 6361 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 6421 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag 6481 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 6541 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 6601 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 6661 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 6721 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 6781 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 6841 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 6901 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 6961 gctcacatgt //