LOCUS       sequence-311846-        7308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of tdTomato fused to LINuS, a light-inducible nuclear
            localization signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.bbrc.2019.11.023"; journalName: "Biochem Biophys Res 
            Commun"; date: "2020-01-29- 29"; volume: "522"; issue: "1"; pages: 
            "138-143"
COMMENT     Sequence Label: pAAV-EF1a-NES-tdTomato-LINuS-WPRE
FEATURES             Location/Qualifiers
     source          1..7308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(5628..5696,5697..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
//
