LOCUS       sequence-402487-        4669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  jAspSnFR3-mRuby3 in Gateway cloning entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4669)
  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 4669)
  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
FEATURES             Location/Qualifiers
     source          1..4669
                     /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..2873
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2898..2997)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3109..3128)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3120..3929
                     /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(3187..3206)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4019..4607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4508..4527
                     /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 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 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 agtcccatca ccatcaccac cattaagaat
     2881 tcgaagctta tcgctagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     2941 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     3001 tccagctgca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     3061 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     3121 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     3181 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     3241 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     3301 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     3361 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     3421 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     3481 cgctggcagt gtccctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     3541 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     3601 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     3661 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     3721 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     3781 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     3841 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     3901 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacattatt
     3961 cagattgggc cccgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4021 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4081 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4141 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4201 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4261 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4321 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4381 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4441 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4501 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4561 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4621 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
