LOCUS       sequence-410599-        7072 bp    DNA     circular SYN 15-APR-2025
DEFINITION  jAspSnFR3-mRuby3 in adenoviral expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7072)
  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 7072)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pAAV-CAG-jAspSnFR3-mRuby3
FEATURES             Location/Qualifiers
     source          1..7072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   61..190
                     /label=AAV2 ITR
     rep_origin      complement(427..1015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(507..526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1186..1977,1978..2046))
                     /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     1809..1828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2047..2151)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2177..2632)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2314..2335)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2526..2545
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2704..2833
                     /label=AAV2 ITR
     primer_bind     3702..3724
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3786..3805
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3839..3848
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4816..4837
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4940..4961)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             6179..6196
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    6219..6807
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6272..6292)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6690..6701)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6850..6971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6861..6880)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6915..6934
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
       61 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
      121 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
      181 gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
      241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
      301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      841 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2161 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2221 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2281 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2341 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2401 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2461 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2521 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2581 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     2641 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     2701 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     2761 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     2821 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
     2881 tctaggaaga tcgtaccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     2941 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     3001 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     3061 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     3121 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
     3181 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
     3241 gcgatggggg cggggggggg gggggggsgs gcgcsggggg gggggggggg gggggggggg
     3301 gggggsgggg sgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
     3361 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
     3421 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
     3481 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
     3541 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
     3601 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggctgtcc
     3661 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     3721 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     3781 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tggatcccgc
     3841 caccatgggc agatctgccg caggcagcac gctggacaaa atcgccaaaa acggtgtgat
     3901 tgtcgtcggt caccgtaaat ctgccgtgcc tttctcttat tacgacagtc agcacaaagt
     3961 ggtgggttac tcgcaggaat actccaacgc cattgttgaa gcagtgaaaa agaaactcaa
     4021 caaaccggac ttgcaggtaa aactgattcc gattacccca caaaaccgta ttccactgct
     4081 gcaaaacggc acttacgatt ttgaatgtgg ttctaccacc aacaacgtcg aacgccaaaa
     4141 acaggcggct ttctctgaca ctattttcgt ggtcggtacg cgcctgttga ccaaaaaggg
     4201 tggcgatatc aaagattttg ccaacctgaa agacaaagcc gtagtcgtca cttccggcac
     4261 tacctctgaa gttttgctca acaaactgaa tgaagagcaa aaaatgaata tgcgcatcat
     4321 cagcgccaaa gatcacggtg actctttccg caccctggaa agcggtcgtg ccgttgcctt
     4381 tatgatggat gacgtgctgc tggccggtga acgtgcgaaa gcgaagaaac cagacgactg
     4441 ggaaatcgtc ggcaagccgc agtctcagga ggcctacggt tgtatgttgc gtaaagatga
     4501 tccgcagttc aaaaagctga tggatgacac catcgctcag gtgcagacct ccggtgaagc
     4561 ggaaaaatgg tttgataagt ggttcaaaaa tccaattctg gtgagccaca tcgtctatat
     4621 caccgccgac aagcagaaga acggcatcaa ggcgaacttc aagatccacc acaacgtgga
     4681 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     4741 cgtgctgctg cccgacaacc actacctgag ctatcagtcc gtgctgagca aagaccccaa
     4801 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccaccacga actctctcgg
     4861 catggacgag ctgtacaagg gcggtaccgg agggagcatg agcaagggcg aggagctgtt
     4921 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     4981 cgtgcgcggc gagggcgagg gcgatgccac caacggcaag ctaaccctga agctcatctg
     5041 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt
     5101 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     5161 gcccgaaggc tacgtccagg agcgcaccat cagcttcaag gacgacggaa cctacaagac
     5221 ccgcgccgag gtgaagttag agggcgacac cctggtgaac cgcatcgagc taaagggcat
     5281 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact ttaacaaccc
     5341 cctgaacatg aatttcgaac tgtcagacga aatgaaagca ctgttcaaag aaccgaatga
     5401 caaggcactg aaactgcagt ctaccggctc cggaaatgct gtgggccagg acacgcagga
     5461 gcgtatggtg tctaagggcg aagagctgat caaggaaaat atgcgtatga aggtggtcat
     5521 ggaaggttcg gtcaacggcc accaattcaa atgcacaggt gaaggagaag gcaggccgta
     5581 cgagggagtg caaaccatga ggatcaaagt catcgaggga ggacccctgc catttgcctt
     5641 tgacattctt gccacgtcgt tcatgtatgg cagccgtact tttatcaagt acccggcaga
     5701 cattcctgat ttctttaaac agtcctttcc tgagggtttt acttgggaaa gagttacgag
     5761 atacgaagat ggtggagtcg tcaccgtcac gcaggacacc agccttgagg atggcgaact
     5821 cgtttacaac gtcaaagtca gaggggtaaa ctttccctcc aatggtcccg tgatgcagaa
     5881 gaagaccaag ggttgggagc ctaatacaga gatgatgtat ccagcagatg gtggtctgag
     5941 gggatacact gatattgcac tgaaagttga tggtggtggc catctgcatt gcaatttcgt
     6001 aacaacttac aggtcaaaaa agaccgtcgg gaacatcaag atgcccggtg tccatgccgt
     6061 tgatcaccgc ctggaaagga tagaggaaag tgacaatgaa acgtacgtag tacaacgcga
     6121 agtcgcagtt gccaagtact ccaatcttgg tggtggtatg gatgaacttt ataagtccca
     6181 tcaccatcac caccattaag aattcgaagc ttatcgataa tcaacctctg gattacaaaa
     6241 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     6301 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     6361 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     6421 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     6481 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     6541 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     6601 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     6661 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     6721 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     6781 ggatctccct ttgggccgcc tccccgcatc gataccgtcg acccgggcgg ccgcttcgag
     6841 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     6901 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     6961 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     7021 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aa
//
