LOCUS       sequence-176804-        7217 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Fluorescent biosensor for super-resolution imaging of ERK activity; 
            targeted to plasma membrane..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7217)
  AUTHORS   Mo GC, Ross B, Hertel F, Manna P, Yang X, Greenwald E, Booth C, 
            Plummer AM, Tenner B, Chen Z, Wang Y, Kennedy EJ, Cole PA, Fleming 
            KG, Palmer A, Jimenez R, Xiao J, Dedecker P, Zhang J
  TITLE     Genetically encoded biosensors for visualizing live-cell biochemical
            activity at super-resolution.
  JOURNAL   Nat Methods. 2017 Apr;14(4):427-434. doi: 10.1038/nmeth.4221. Epub 
            2017 Mar 13.
  PUBMED    28288122
REFERENCE   2  (bases 1 to 7217)
  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"; doi: 
            "10.1038/nmeth.4221"; journalName: "Nat Methods"; date: "2017-04"; 
            volume: "14"; issue: "4"; pages: "427-434"
COMMENT     Sequence Label: Lyn-FLINC-EKAR1
FEATURES             Location/Qualifiers
     source          1..7217
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             217..912
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTDMALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHK"
     promoter        complement(973..991)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(974..991)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1011..1028)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1017..1241
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1287..1715
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1374..1393)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1584..1605
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1724..1744)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1729..2058
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1909..2044
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1971..1990
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2125..2919
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2179..2198)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2789..2808
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3093..3214
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3130..3149)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3184..3203
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3263..3279)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3263..3279)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3276..3298)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3287..3303
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3311..3317,3318..3335,3336..3341))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3356..3377
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3494..3511)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3665..4253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3745..3764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4424..5215,5216..5284))
                     /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     5047..5066
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5285..5389)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5464..5483)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5655..6034
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6035..6238
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6189..6209
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6283..6302
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6283..6301
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6321..6330
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6366..7037
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
ORIGIN
        1 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggttccggc
       61 agtgctggtg gtagtgctgg tggtagtacc agtgctggtg gtagtgctgg tggtagtgct
      121 ggtggtagtg ctggtggtag tgctggtggt tccggaccag atgtccctag aactccagtg
      181 gataaagcaa agctgtcatt ccaatttccg ggtaccatga gcgagctgat taaggagaac
      241 atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa gtgcacatcc
      301 gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc
      361 ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg cagcagaacc
      421 ttcatcaacc acacccaggg catccccgat ttctttaagc agtccttccc tgagggcttc
      481 acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc
      541 agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa cttcccatcc
      601 aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccaacaccga gatgctgtac
      661 cccgctgacg gcggcctgga aggcagaacc gacatggccc tgaagctcgt gggcgggggc
      721 cacctgatct gcaacttcaa gaccacatac agatccaaga aacccgctaa gaacctcaag
      781 atgcccggcg tctactatgt ggaccacaga ctggaaagaa tcaaggaggc cgacaaagag
      841 acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc tagcaaactg
      901 gggcacaagt aagaattctg cagatatcca tcacactggc ggccgctcga gcatgcatct
      961 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
     1021 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1081 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1141 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1201 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     1261 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     1321 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1381 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1441 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1501 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1561 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1621 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1681 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     1741 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     1801 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     1861 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     1921 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     1981 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2041 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     2101 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     2161 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2221 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2281 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     2341 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     2401 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     2461 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     2521 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     2581 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     2641 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     2701 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     2761 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     2821 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     2881 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     2941 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     3001 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3061 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3121 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3181 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     3241 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3301 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3361 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3421 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3481 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3541 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3601 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3661 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3721 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3781 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3841 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3901 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3961 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4021 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4081 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4141 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4201 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4261 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4321 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4381 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4441 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4501 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4561 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4621 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4681 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4741 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4801 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4861 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4921 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4981 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5041 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5101 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5161 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5221 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5281 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5341 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5401 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     5461 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     5521 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     5581 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     5641 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     5701 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     5761 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     5821 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     5881 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     5941 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     6001 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     6061 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     6121 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     6181 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
     6241 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
     6301 ggagacccaa gcttgcggcc gccaccatgg gctgcatcaa gagcaagcgc aaggacaagg
     6361 atccgatgag tgtgattaaa ccagacatga agatcaagct gcgtatggaa ggcgctgtaa
     6421 atggacaccc gttcgcgatt gaaggagttg gccttgggaa gcctttcgag ggaaaacaga
     6481 gtatggacct taaagtcaaa gaaggcggac ctctgccttt cgcctatgac atcttgacaa
     6541 ctgtgttctg ttacggcaac agggtattcg ccaaataccc agaaaatata gtagactatt
     6601 tcaagcagtc gtttcctgag ggctactctt gggaacgaag catgaattac gaagacgggg
     6661 gcatttgtaa cgcgacaaac gacataaccc tggatggtga ctgttatatc tatgaaattc
     6721 gatttgatgg tgtgaacttt cctgccaatg gtccagttat gcagaagagg actgtgaaat
     6781 gggagccatc cactgagaaa ttgtatgtgc gtgatggagt gctgaagggt gatgttaaca
     6841 tggctctgtc gcttgaagga ggtggccatt accgatgtga cttcaaaact acttataaag
     6901 ctaagaaggt tgtccagttg ccagactatc actttgtgga ccaccacatt gagattaaaa
     6961 gccacgacaa agattacagt aatgttaatc tgcatgagca tgccgaagcg cattctgagc
     7021 tgccgaggca ggccaagaga tctatggcgg acgaggagaa gctgccgccc ggctgggaga
     7081 agcgcatgag ccgcagctca ggccgagtgt actacttcaa ccacatcact aacgccagcc
     7141 agtgggagcg gcccagcggc aacagcagca gtggtggcaa aaacgggcag ggggagcctg
     7201 ccaggggtac cagtgct
//
