LOCUS Exported 7308 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION AAV expression of tdTomato fused to LINuS, a light-inducible nuclear localization signal. ACCESSION . VERSION . KEYWORDS pAAV-EF1a-NES-tdTomato-LINuS-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7308) AUTHORS Inutsuka A, Kimizuka N, Takanohashi N, Yakabu H, Onaka T TITLE Visualization of a blue light transmission area in living animals using light-induced nuclear translocation of fluorescent proteins. JOURNAL Biochem Biophys Res Commun. 2020 Jan 29;522(1):138-143. doi: 10.1016/j.bbrc.2019.11.023. Epub 2019 Nov 19. PUBMED 31757418 REFERENCE 2 (bases 1 to 7308) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7308 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 231..1409 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 462..1400 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1357..1377 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 1467..2894 /codon_start=1 /product="tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)" /label=tdTomato /note="mammalian codon-optimized" /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE RSEGRHHLFLYGMDELYK" misc_feature 3435..4023 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3488..3508) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3906..3917) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4055..4531 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4254..4273) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4571..4711 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4571..4700 /label=AAV2 ITR rep_origin 4786..5241 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4873..4892) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5083..5104 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5258..5277) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5377..5399 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5437..5455) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5523..5627 /gene="bla" /label=AmpR promoter CDS 5628..6488 /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(5846..5865) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6659..7247 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7148..7167 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga 181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg 241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg 301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt 361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg 421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt 481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct 541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga 601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct 661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt 721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc 781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg 841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac 901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc 961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag 1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct 1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg 1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga 1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc 1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg 1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa 1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc cgccaccatg ttagccttga 1441 aattagcagg tcttgatatc gctagcatgg tgagcaaggg cgaggaggtc atcaaagagt 1501 tcatgcgctt caaggtgcgc atggagggct ccatgaacgg ccacgagttc gagatcgagg 1561 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg 1621 gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcatgtac ggctccaagg 1681 cgtacgtgaa gcaccccgcc gacatccccg attacaagaa gctgtccttc cccgagggct 1741 tcaagtggga gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg acccaggact 1801 cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc 1861 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt 1921 acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg aaggacggcg 1981 gccactacct ggtggagttc aagaccatct acatggccaa gaagcccgtg caactgcccg 2041 gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg 2101 tggaacagta cgagcgctcc gagggccgcc accacctgtt cctggggcat ggcaccggca 2161 gcaccggcag cggcagctcc ggcaccgcct cctccgagga caacaacatg gccgtcatca 2221 aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga 2281 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga 2341 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct 2401 ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg 2461 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc 2521 aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact 2581 tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc 2641 gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg 2701 acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac 2761 tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca 2821 ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg tacggcatgg 2881 acgagctgta caagggtgga tctggaggtt caggtggaag tttggctact acacttgaac 2941 gtattgagaa gaactttgtc attactgacc caagattgcc agataatccc attatattcg 3001 cgtccgatag tttcttgcag ttgacagaat atagccgtga agaaattttg ggaagaaact 3061 gcaggtttct acaaggtcct gaaactgatc gcgcgacagt gagaaaaatt agagatgcca 3121 tagataacca aacagaggtc actgttcagc tgattaatta tacaaagagt ggtaaaaagt 3181 tctggaacct ctttcacttg cagcctatgc gagatcagaa gggagatgtc cagtacttta 3241 ttggggttca gttggatgga actgagcatg tccgagatgc tgccgagaga gagggagtca 3301 tgctgattaa gaaaactgca gaaaatattg acgaggccgc aaagagactg cccgacgcca 3361 acctggccgc agcagccgca gccgcagcaa agaagaaaaa gctggactag gaattcgata 3421 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc 3481 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg 3541 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc 3601 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg 3661 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg 3721 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga 3781 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct 3841 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg 3901 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc 3961 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc 4021 cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc 4081 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa 4141 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg 4201 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg 4261 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt 4321 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag 4381 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt 4441 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca 4501 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc 4561 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg 4621 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca 4681 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg 4741 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg 4801 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg 4861 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc 4921 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc 4981 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga 5041 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa 5101 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga 5161 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 5221 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat 5281 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc 5341 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt 5401 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat 5461 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg 5521 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga 5581 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac 5641 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 5701 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca 5761 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc 5821 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 5881 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac 5941 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 6001 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg 6061 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 6121 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 6181 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat 6241 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 6301 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 6361 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 6421 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 6481 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt 6541 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt 6601 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 6661 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 6721 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 6781 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca 6841 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 6901 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 6961 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 7021 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 7081 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 7141 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 7201 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 7261 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt //