LOCUS       sequence-402491-        4661 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nuclear directed jAspSnFR3-mRuby3 in Gateway cloning entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4661)
  AUTHORS   Davidsen K, Marvin JS, Aggarwal A, Brown TA, Sullivan LB
  TITLE     An engineered biosensor enables dynamic aspartate measurements in 
            living cells.
  JOURNAL   eLife. 2024 Feb 23;12:RP90024. doi: 10.7554/eLife.90024.
  PUBMED    38393319
REFERENCE   2  (bases 1 to 4661)
  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"; journalName: "eLife."; 
            date: "2024-02-23"; volume: "12:RP90024. doi"; pages: " 
            10.7554/eLife.90024"
COMMENT     Sequence Label: pENTR1a-jAspSnFR3-mRuby3-NLS
FEATURES             Location/Qualifiers
     source          1..4661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      516..525
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1493..1514
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1617..1638)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2856..2882
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     protein_bind    complement(2890..2989)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3101..3120)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3112..3921
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3179..3198)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4011..4599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4500..4519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaggc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac
      481 tggatccggt accgaattcg cttactaaaa gccaggccac catgggcaga tctgccgcag
      541 gcagcacgct ggacaaaatc gccaaaaacg gtgtgattgt cgtcggtcac cgtaaatctg
      601 ccgtgccttt ctcttattac gacagtcagc acaaagtggt gggttactcg caggaatact
      661 ccaacgccat tgttgaagca gtgaaaaaga aactcaacaa accggacttg caggtaaaac
      721 tgattccgat taccccacaa aaccgtattc cactgctgca aaacggcact tacgattttg
      781 aatgtggttc taccaccaac aacgtcgaac gccaaaaaca ggcggctttc tctgacacta
      841 ttttcgtggt cggtacgcgc ctgttgacca aaaagggtgg cgatatcaaa gattttgcca
      901 acctgaaaga caaagccgta gtcgtcactt ccggcactac ctctgaagtt ttgctcaaca
      961 aactgaatga agagcaaaaa atgaatatgc gcatcatcag cgccaaagat cacggtgact
     1021 ctttccgcac cctggaaagc ggtcgtgccg ttgcctttat gatggatgac gtgctgctgg
     1081 ccggtgaacg tgcgaaagcg aagaaaccag acgactggga aatcgtcggc aagccgcagt
     1141 ctcaggaggc ctacggttgt atgttgcgta aagatgatcc gcagttcaaa aagctgatgg
     1201 atgacaccat cgctcaggtg cagacctccg gtgaagcgga aaaatggttt gataagtggt
     1261 tcaaaaatcc aattctggtg agccacatcg tctatatcac cgccgacaag cagaagaacg
     1321 gcatcaaggc gaacttcaag atccaccaca acgtggagga cggcagcgtg cagctcgccg
     1381 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1441 acctgagcta tcagtccgtg ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     1501 tgctggagtt cgtgaccgcc accacgaact ctctcggcat ggacgagctg tacaagggcg
     1561 gtaccggagg gagcatgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1621 tcgagctgga cggcgacgta aacggccaca agttcagcgt gcgcggcgag ggcgagggcg
     1681 atgccaccaa cggcaagcta accctgaagc tcatctgcac caccggcaag ctgcccgtgc
     1741 cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg
     1801 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1861 gcaccatcag cttcaaggac gacggaacct acaagacccg cgccgaggtg aagttagagg
     1921 gcgacaccct ggtgaaccgc atcgagctaa agggcatcga cttcaaggag gacggcaaca
     1981 tcctggggca caagctggag tacaacttta acaaccccct gaacatgaat ttcgaactgt
     2041 cagacgaaat gaaagcactg ttcaaagaac cgaatgacaa ggcactgaaa ctgcagtcta
     2101 ccggctccgg aaatgctgtg ggccaggaca cgcaggagcg tatggtgtct aagggcgaag
     2161 agctgatcaa ggaaaatatg cgtatgaagg tggtcatgga aggttcggtc aacggccacc
     2221 aattcaaatg cacaggtgaa ggagaaggca ggccgtacga gggagtgcaa accatgagga
     2281 tcaaagtcat cgagggagga cccctgccat ttgcctttga cattcttgcc acgtcgttca
     2341 tgtatggcag ccgtactttt atcaagtacc cggcagacat tcctgatttc tttaaacagt
     2401 cctttcctga gggttttact tgggaaagag ttacgagata cgaagatggt ggagtcgtca
     2461 ccgtcacgca ggacaccagc cttgaggatg gcgaactcgt ttacaacgtc aaagtcagag
     2521 gggtaaactt tccctccaat ggtcccgtga tgcagaagaa gaccaagggt tgggagccta
     2581 atacagagat gatgtatcca gcagatggtg gtctgagggg atacactgat attgcactga
     2641 aagttgatgg tggtggccat ctgcattgca atttcgtaac aacttacagg tcaaaaaaga
     2701 ccgtcgggaa catcaagatg cccggtgtcc atgccgttga tcaccgcctg gaaaggatag
     2761 aggaaagtga caatgaaacg tacgtagtac aacgcgaagt cgcagttgcc aagtactcca
     2821 atcttggtgg tggtatggat gaactttata agtcccctgc tgcaaagcga gtgaagctcg
     2881 attaactaga cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     2941 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     3001 cagctctggc ccgtgtctca aaatctctga tgttacatag cacaagataa aaatatatca
     3061 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     3121 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     3181 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     3241 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     3301 acagatgaga tggtcagact aaactggctg acggaattta tgccccttcc gaccatcaag
     3361 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     3421 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     3481 gtgtccctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     3541 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     3601 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     3661 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     3721 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     3781 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     3841 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     3901 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacatta ttcagattgg
     3961 gccccgttcc actgagcgtc agaccccgta gaaaagatca aaggatcctc ttgagatcct
     4021 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4081 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4141 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4201 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4261 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4321 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4381 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4441 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4501 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4561 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4621 ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
