LOCUS Exported 7088 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV-mediated expression of Chronos-GFP under the EF1-alpha promoter (1.1kb short version), in floxed/reversed (Cre-dependent) manner. Using bGHpA signal.. ACCESSION . VERSION . KEYWORDS pAAV-EF1-alpha-1.1-FLEX-rc [Chronos-GFP] SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7088) AUTHORS Klapoetke NC, Murata Y, Kim SS, Pulver SR, Birdsey-Benson A, Cho YK, Morimoto TK, Chuong AS, Carpenter EJ, Tian Z, Wang J, Xie Y, Yan Z, Zhang Y, Chow BY, Surek B, Melkonian M, Jayaraman V, Constantine-Paton M, Wong GK, Boyden ES TITLE Independent optical excitation of distinct neural populations. JOURNAL Nat Methods. 2014 Mar;11(3):338-46. doi: 10.1038/nmeth.2836. Epub 2014 Feb 9. PUBMED 24509633 REFERENCE 2 (bases 1 to 7088) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7088 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(11..28) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(182..770) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(262..281) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(941..1801) /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 1564..1583 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1802..1906) /gene="bla" /label=AmpR promoter rep_origin complement(1932..2387) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2069..2090) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2281..2300 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 2459..2588 /label=AAV2 ITR promoter 2661..3839 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2892..3830 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3787..3807 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind complement(3848..3881) /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(3944..3977) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(3994..4707) /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(4036..4057) /label=EGFP-C /note="EGFP, forward primer" primer_bind 4385..4404 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4644..4665 /label=EGFP-N /note="EGFP, reverse primer" regulatory 5697..5706 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind 5728..5761 /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 5832..5865 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 5898..6486 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5951..5971) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6369..6380) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 6527..6734 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 6904..7033 /label=AAV2 ITR ORIGIN 1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta 1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga 2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa 2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga 2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc 2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga 2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg 2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt 2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct 2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag 2641 gaagatcgta ggttaattaa ggctccggtg cccgtcagtg ggcagagcgc acatcgccca 2701 cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc 2761 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg 2821 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg 2881 ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg 2941 gcccttgcgt gccttgaatt acttccacct ggctgcagta cgtgattctt gatcccgagc 3001 ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc 3061 gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc 3121 ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg 3181 ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg 3241 gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt 3301 cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg 3361 gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa 3421 ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg 3481 cagggagctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac 3541 aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg 3601 cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg 3661 gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc 3721 cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt 3781 tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag 3841 aattcccata acttcgtata aagtatccta tacgaagtta tatcaaaata ggaagaccaa 3901 tgcttcacca tcgacccgaa ttgccaagca tcaccatcga cccataactt cgtataatgt 3961 atgctatacg aagttatact agctagctca ttacttgtac agctcgtcca tgccgagagt 4021 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc 4081 tttgctcagg gcggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc 4141 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt 4201 gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac 4261 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa 4321 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc 4381 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc 4441 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca 4501 ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt 4561 ggtgcaaatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga 4621 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc 4681 ggtgaacagc tcctcgccct tgctcactgc tactaccggt gccgccactc ctccctcctc 4741 ctcctcgtcg acgaaggtct cgatctccat gttctcgccg gccacgttga tggtggttgt 4801 cttccggatg tcgccgtgga tcaggatgtg ctcgtggatc ttcactctca ggtggtggcc 4861 caggaagccc caagtgttct tgctgatcag atccaggatc aggtggccga ttccgctggt 4921 gtaaggtgtg atcaggccca gtccctcgtg tccggcgatg aacatcacgg ggaacatcag 4981 ccaagagcag aagaagacgt aggccatgat cttgcaaatc ttccggcaga cgcctttggg 5041 cagggtgtgg tagctctcga tatacacctt ggcgatctgg aagaagcaag tcacgccgta 5101 gaacaggccg atcatgaaga acaggatctt cagggggccc tttgtaaagg cggctgtgat 5161 cccccacacg atgttgccga tgtcggtcac caggatggtc atggtccgct tgctgtactc 5221 ttcgtgcagt ccggtcaggt tgctcaggtg gatcaggatc acggggcaag tcaggagcca 5281 catgctgtac cgcagccaaa tcacggctcc tccgttggtc tggtacactg tggctgggct 5341 gtcgacctcg tggaacagct cgatgaagca cttgacgagc tcgatcacgc agacgtagac 5401 ctcctcccag ccggttgtgg ccttgaagct gtgccaaccg tagaagatga gctgcacgat 5461 agccacgatc acggtgaacc actggagtcc cactgcgatc ttgtgctgca gctcggttcc 5521 gtggttgatg tgggggaagc aaccgtgatc ggctccggct gttgtggcgt tagaggtttc 5581 tccgtgagcg tctgctgctg gtgtcaccac tgctgcggcg ctcagcaggc ctctaattgt 5641 ggcttcggcg ctaggcacgg ctgagataaa ggcgtgggtc attgtggcgg ctgtttccat 5701 ggtggcggcg cggcctaggt acctagtata acttcgtata ggatacttta tacgaagtta 5761 tcattgggat tcttcctatt ttgatccaag catcaccatc gaccctctag tccagatctc 5821 accatcgacc cataacttcg tatagcatac attatacgaa gttatgtccc tcgaagaggt 5881 tcgcgaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta 5941 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc 6001 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt 6061 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg 6121 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt 6181 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct 6241 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt 6301 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct 6361 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc 6421 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct 6481 ccccgcatcg ataccgtcga cccgggcggc cgctcataaa gctcgactgt gccttctagt 6541 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact 6601 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat 6661 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc 6721 aggcatgctg gggatgcggt ccggactgta caattgcatt cattttatgt ttcaggttca 6781 gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga 6841 taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact 6901 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg 6961 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg 7021 agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 7081 tgggcgct //