LOCUS       sequence-271857-        6698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Salsa6f is a fluorescent ratiometric genetically encoded calcium 
            indicator, (tdTomato linked to GCaMP6f by a V5 epitope tag).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6698)
  AUTHORS   Dong TX, Othy S, Jairaman A, Skupsky J, Zavala A, Parker I, Dynes 
            JL, Cahalan MD
  TITLE     T-cell calcium dynamics visualized in a ratiometric tdTomato-GCaMP6f
            transgenic reporter mouse.
  JOURNAL   eLife. 2017 Dec 14;6. pii: 32417. doi: 10.7554/eLife.32417.
  PUBMED    29239725
REFERENCE   2  (bases 1 to 6698)
  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: "2017-12-14"; volume: "6. pii: 32417. doi"; pages: " 
            10.7554/eLife.32417"
COMMENT     Sequence Label: Salsa6f
FEATURES             Location/Qualifiers
     source          1..6698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..2040
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             2050..2091
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      2104..2113
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2125..2181
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     2394..2415
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2521..2542)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2782..2801)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3486..3607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3523..3542)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3577..3596
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3614..4069)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3751..3772)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3963..3982
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4096..4200
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4202..4559
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4202..4393
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4225..4245)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4410..4545
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4594..5388
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4594..5388
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4648..4667)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5258..5277
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5576..5595)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5620..5667
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5996..6584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6485..6504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag
      661 gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
      721 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc
      781 ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac
      841 cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc
      901 gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac
      961 ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta
     1021 atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc
     1081 gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg
     1141 gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg
     1201 gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag
     1261 cgctccgagg gccgccacca cctgttcctg gggcatggca ccggcagcac cggcagcggc
     1321 agctccggca ccgcctcctc cgaggacaac aacatggccg tcatcaaaga gttcatgcgc
     1381 ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc
     1441 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     1501 ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg
     1561 aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg
     1621 gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg
     1681 caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc
     1741 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc
     1801 gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac
     1861 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac
     1921 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1981 tacgagcgct ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag
     2041 cagatctcgg gtaagcctat ccctaaccct ctcctcggtc tcgattctac ggatccaccg
     2101 gtcgccacca tggtcgactc atcacgtcgt aagtggaata agacaggtca cgcagtcaga
     2161 gctataggtc ggctgagctc actcgagaac gtctatatca aggccgacaa gcagaagaac
     2221 ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     2281 taccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2341 tacctgagcg tgcagtccaa actttcgaaa gaccccaacg agaagcgcga tcacatggtc
     2401 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc
     2461 ggtaccggag ggagcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     2521 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggtgag
     2581 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     2641 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     2701 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacatccag
     2761 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     2821 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2881 aacatcctgg ggcacaagct ggagtacaac ctgccggacc aactgactga agagcagatc
     2941 gcagaattta aagaggaatt ctccctattt gacaaggacg gggatgggac aataacaacc
     3001 aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc agagctgcag
     3061 gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc tgagttcctg
     3121 acaatgatgg caagaaaaat gaaatacagg gacacggaag aagaaattag agaagcgttc
     3181 ggtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg ccacgtgatg
     3241 acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag ggaagcagac
     3301 atcgatgggg atggtcaggt aaactacgaa gagtttgtac aaatgatgac agcgaagctt
     3361 taaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     3421 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3481 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3541 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3601 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3661 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3721 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3781 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3841 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3901 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3961 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4021 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     4081 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4141 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4201 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4261 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4321 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4381 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4441 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4501 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     4561 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4621 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4681 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4741 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     4801 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4861 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4921 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4981 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5041 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5101 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     5161 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5221 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5281 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5341 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5401 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5461 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5521 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     5581 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     5641 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     5701 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     5761 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     5821 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     5881 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5941 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6001 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6061 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6121 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     6181 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6241 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6301 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6361 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6421 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6481 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6541 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6601 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6661 cccctgattc tgtggataac cgtattaccg ccatgcat
//
