LOCUS       sequence-402490-        5023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mitochondrial directed jAspSnFR3-mRuby3 in Gateway cloning entry 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5023)
  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 5023)
  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-mRuby3
FEATURES             Location/Qualifiers
     source          1..5023
                     /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     1847..1868
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1971..1992)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3210..3227
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(3252..3351)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3463..3482)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3474..4283
                     /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(3541..3560)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4373..4961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4862..4881
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 tgacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc 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 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 cagatctgcc gcaggcagca
      901 cgctggacaa aatcgccaaa aacggtgtga ttgtcgtcgg tcaccgtaaa tctgccgtgc
      961 ctttctctta ttacgacagt cagcacaaag tggtgggtta ctcgcaggaa tactccaacg
     1021 ccattgttga agcagtgaaa aagaaactca acaaaccgga cttgcaggta aaactgattc
     1081 cgattacccc acaaaaccgt attccactgc tgcaaaacgg cacttacgat tttgaatgtg
     1141 gttctaccac caacaacgtc gaacgccaaa aacaggcggc tttctctgac actattttcg
     1201 tggtcggtac gcgcctgttg accaaaaagg gtggcgatat caaagatttt gccaacctga
     1261 aagacaaagc cgtagtcgtc acttccggca ctacctctga agttttgctc aacaaactga
     1321 atgaagagca aaaaatgaat atgcgcatca tcagcgccaa agatcacggt gactctttcc
     1381 gcaccctgga aagcggtcgt gccgttgcct ttatgatgga tgacgtgctg ctggccggtg
     1441 aacgtgcgaa agcgaagaaa ccagacgact gggaaatcgt cggcaagccg cagtctcagg
     1501 aggcctacgg ttgtatgttg cgtaaagatg atccgcagtt caaaaagctg atggatgaca
     1561 ccatcgctca ggtgcagacc tccggtgaag cggaaaaatg gtttgataag tggttcaaaa
     1621 atccaattct ggtgagccac atcgtctata tcaccgccga caagcagaag aacggcatca
     1681 aggcgaactt caagatccac cacaacgtgg aggacggcag cgtgcagctc gccgaccact
     1741 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1801 gctatcagtc cgtgctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1861 agttcgtgac cgccaccacg aactctctcg gcatggacga gctgtacaag ggcggtaccg
     1921 gagggagcat gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1981 tggacggcga cgtaaacggc cacaagttca gcgtgcgcgg cgagggcgag ggcgatgcca
     2041 ccaacggcaa gctaaccctg aagctcatct gcaccaccgg caagctgccc gtgccctggc
     2101 ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac cccgaccaca
     2161 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2221 tcagcttcaa ggacgacgga acctacaaga cccgcgccga ggtgaagtta gagggcgaca
     2281 ccctggtgaa ccgcatcgag ctaaagggca tcgacttcaa ggaggacggc aacatcctgg
     2341 ggcacaagct ggagtacaac tttaacaacc ccctgaacat gaatttcgaa ctgtcagacg
     2401 aaatgaaagc actgttcaaa gaaccgaatg acaaggcact gaaactgcag tctaccggct
     2461 ccggaaatgc tgtgggccag gacacgcagg agcgtatggt gtctaagggc gaagagctga
     2521 tcaaggaaaa tatgcgtatg aaggtggtca tggaaggttc ggtcaacggc caccaattca
     2581 aatgcacagg tgaaggagaa ggcaggccgt acgagggagt gcaaaccatg aggatcaaag
     2641 tcatcgaggg aggacccctg ccatttgcct ttgacattct tgccacgtcg ttcatgtatg
     2701 gcagccgtac ttttatcaag tacccggcag acattcctga tttctttaaa cagtcctttc
     2761 ctgagggttt tacttgggaa agagttacga gatacgaaga tggtggagtc gtcaccgtca
     2821 cgcaggacac cagccttgag gatggcgaac tcgtttacaa cgtcaaagtc agaggggtaa
     2881 actttccctc caatggtccc gtgatgcaga agaagaccaa gggttgggag cctaatacag
     2941 agatgatgta tccagcagat ggtggtctga ggggatacac tgatattgca ctgaaagttg
     3001 atggtggtgg ccatctgcat tgcaatttcg taacaactta caggtcaaaa aagaccgtcg
     3061 ggaacatcaa gatgcccggt gtccatgccg ttgatcaccg cctggaaagg atagaggaaa
     3121 gtgacaatga aacgtacgta gtacaacgcg aagtcgcagt tgccaagtac tccaatcttg
     3181 gtggtggtat ggatgaactt tataagtccc atcaccatca ccaccattaa gaattcgaag
     3241 cttatcgcta gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     3301 aatttgttgc gacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     3361 tgcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     3421 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     3481 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     3541 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     3601 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     3661 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     3721 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     3781 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     3841 cagtgtccct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     3901 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     3961 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     4021 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     4081 tttttgacga ggggaaatta ataggttgta ttgatgttgg gcgagtcgga atcgcagacc
     4141 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     4201 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     4261 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacat tattcagatt
     4321 gggccccgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4381 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4441 tttgtttgcc ggatcaagag ctaccaactc tttttccgag ggtaactggc ttcagcagag
     4501 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4561 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4621 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4681 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4741 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4801 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4861 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4921 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4981 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
