LOCUS       sequence-402515-        4243 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mitochondrial directed jAspSnFR3 in Gateway cloning entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4243)
  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 4243)
  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-MTS-jAspSnFR3
FEATURES             Location/Qualifiers
     source          1..4243
                     /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     1844..1865
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1968..1989)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(2472..2571)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2683..2702)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2694..3503
                     /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(2761..2780)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3593..4181
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4082..4101
                     /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 gctgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccga
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac
      481 tggatccggt accgaattcg cttactaaaa gccaggccac catggcactg cttcgcgccg
      541 ccgtttcaga actcagacgg agaggacggg gtgcgcttac tcccctcccg gcgctgtcta
      601 gcttgctttc ctcacttagc ccccgaagtc ccgcctcaac gcgcccagag ccaaacaatc
      661 cacacgcaga tcgacgccat gtcatcgctt tgaggcgatg ccccccactt cctgcctctg
      721 ccgttctggc acctgaactc ctgcatgcac gaggattgct cccgagacat tggtctcatg
      781 cctctccctt gtccacgtcc tcttcatcca gtagaccagc agataaggcg cagttgacct
      841 gggtcgataa atggatccca gaagccgcga gaccgtatgg atccgccgca ggcagcacgc
      901 tggacaaaat cgccaaaaac ggtgtgattg tcgtcggtca ccgtaaatct gccgtgcctt
      961 tctcttatta cgacagtcag cacaaagtgg tgggttactc gcaggaatac tccaacgcca
     1021 ttgttgaagc agtgaaaaag aaactcaaca aaccggactt gcaggtaaaa ctgattccga
     1081 ttaccccaca aaaccgtatt ccactgctgc aaaacggcac ttacgatttt gaatgtggtt
     1141 ctaccaccaa caacgtcgaa cgccaaaaac aggcggcttt ctctgacact attttcgtgg
     1201 tcggtacgcg cctgttgacc aaaaagggtg gcgatatcaa agattttgcc aacctgaaag
     1261 acaaagccgt agtcgtcact tccggcacta cctctgaagt tttgctcaac aaactgaatg
     1321 aagagcaaaa aatgaatatg cgcatcatca gcgccaaaga tcacggtgac tctttccgca
     1381 ccctggaaag cggtcgtgcc gttgccttta tgatggatga cgtgctgctg gccggtgaac
     1441 gtgcgaaagc gaagaaacca gacgactggg aaatcgtcgg caagccgcag tctcaggagg
     1501 cctacggttg tatgttgcgt aaagatgatc cgcagttcaa aaagctgatg gatgacacca
     1561 tcgctcaggt gcagacctcc ggtgaagcgg aaaaatggtt tgataagtgg ttcaaaaatc
     1621 caattctggt gagccacatc gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1681 cgaacttcaa gatccaccac aacgtggagg acggcagcgt gcagctcgcc gaccactacc
     1741 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1801 atcagtccgt gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1861 tcgtgaccgc caccacgaac tctctcggca tggacgagct gtacaagggc ggtaccggag
     1921 ggagcatgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1981 acggcgacgt aaacggccac aagttcagcg tgcgcggcga gggcgagggc gatgccacca
     2041 acggcaagct aaccctgaag ctcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2101 ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga
     2161 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatca
     2221 gcttcaagga cgacggaacc tacaagaccc gcgccgaggt gaagttagag ggcgacaccc
     2281 tggtgaaccg catcgagcta aagggcatcg acttcaagga ggacggcaac atcctggggc
     2341 acaagctgga gtacaacttt aacaaccccc tgaacatgaa tttcgaactg tcagacgaaa
     2401 tgaaagcact gttcaaagaa ccgaatgaca aggcactgaa ataagaattc gaagcttgat
     2461 ccggctgcta gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2521 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2581 tgcagctctg gcccgtgtct caaaatctct gatgttacac tgcacaagat aaaaatatat
     2641 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     2701 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     2761 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     2821 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     2881 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     2941 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     3001 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     3061 cagtgtccct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     3121 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     3181 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     3241 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     3301 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     3361 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     3421 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     3481 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacat tattcagatt
     3541 gggccccgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3601 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3661 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3721 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3781 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3841 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3901 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3961 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4021 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4081 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4141 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4201 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
