LOCUS       sequence-176873-        7217 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Negative-control mutant of Lyn-FLINC-EKAR1..
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(TA)
FEATURES             Location/Qualifiers
     source          1..7217
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             216..911
                     /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(972..990)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(973..990)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1010..1027)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1016..1240
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1286..1714
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1373..1392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1583..1604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1723..1743)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1728..2057
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1908..2043
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1970..1989
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2124..2918
                     /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(2178..2197)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2788..2807
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3092..3213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3129..3148)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3183..3202
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3262..3278)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3262..3278)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3275..3297)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3286..3302
                     /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(3310..3316,3317..3334,3335..3340))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3355..3376
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3493..3510)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3664..4252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3744..3763)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4423..5214,5215..5283))
                     /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     5046..5065
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5284..5388)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5463..5482)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5654..6033
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6034..6237
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6188..6208
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6282..6301
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6282..6300
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6320..6329
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6365..7036
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
ORIGIN
        1 gtggtagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggttccggca
       61 gtgctggtgg tagtgctggt ggtagtacca gtgctggtgg tagtgctggt ggtagtgctg
      121 gtggtagtgc tggtggtagt gctggtggtt ccggaccaga tgtccctaga gccccagtgg
      181 ataaagcaaa gctgtcattc caatttccgg gtaccatgag cgagctgatt aaggagaaca
      241 tgcacatgaa gctgtacatg gagggcaccg tgaacaacca ccacttcaag tgcacatccg
      301 agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg
      361 gccctctccc cttcgccttc gacatcctgg ctaccagctt catgtacggc agcagaacct
      421 tcatcaacca cacccagggc atccccgatt tctttaagca gtccttccct gagggcttca
      481 catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca
      541 gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcccatcca
      601 acggccctgt gatgcagaag aaaacactcg gctgggaggc caacaccgag atgctgtacc
      661 ccgctgacgg cggcctggaa ggcagaaccg acatggccct gaagctcgtg ggcgggggcc
      721 acctgatctg caacttcaag accacataca gatccaagaa acccgctaag aacctcaaga
      781 tgcccggcgt ctactatgtg gaccacagac tggaaagaat caaggaggcc gacaaagaga
      841 cctacgtcga gcagcacgag gtggctgtgg ccagatactg cgacctccct agcaaactgg
      901 ggcacaagta agaattctgc agatatccat cacactggcg gccgctcgag catgcatcta
      961 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     1021 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1081 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1141 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     1201 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     1261 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1321 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1381 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1441 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1501 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1561 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1621 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1681 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     1741 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     1801 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     1861 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     1921 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     1981 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2041 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     2101 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     2161 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     2221 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     2281 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     2341 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     2401 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     2461 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     2521 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     2581 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     2641 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     2701 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     2761 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     2821 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     2881 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     2941 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     3001 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3061 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     3121 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3181 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     3241 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3301 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3361 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3421 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3481 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3541 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3601 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3661 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3721 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3781 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3841 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3901 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3961 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4021 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4081 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4141 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4201 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4261 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4321 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4381 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4441 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4501 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4561 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4621 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4681 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4741 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4801 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4861 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4921 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4981 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5041 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5101 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5161 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5221 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5281 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5341 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5401 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
     5461 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     5521 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     5581 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     5641 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     5701 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     5761 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     5821 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     5881 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     5941 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     6001 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     6061 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     6121 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     6181 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
     6241 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
     6301 gagacccaag cttgcggccg ccaccatggg ctgcatcaag agcaagcgca aggacaagga
     6361 tccgatgagt gtgattaaac cagacatgaa gatcaagctg cgtatggaag gcgctgtaaa
     6421 tggacacccg ttcgcgattg aaggagttgg ccttgggaag cctttcgagg gaaaacagag
     6481 tatggacctt aaagtcaaag aaggcggacc tctgcctttc gcctatgaca tcttgacaac
     6541 tgtgttctgt tacggcaaca gggtattcgc caaataccca gaaaatatag tagactattt
     6601 caagcagtcg tttcctgagg gctactcttg ggaacgaagc atgaattacg aagacggggg
     6661 catttgtaac gcgacaaacg acataaccct ggatggtgac tgttatatct atgaaattcg
     6721 atttgatggt gtgaactttc ctgccaatgg tccagttatg cagaagagga ctgtgaaatg
     6781 ggagccatcc actgagaaat tgtatgtgcg tgatggagtg ctgaagggtg atgttaacat
     6841 ggctctgtcg cttgaaggag gtggccatta ccgatgtgac ttcaaaacta cttataaagc
     6901 taagaaggtt gtccagttgc cagactatca ctttgtggac caccacattg agattaaaag
     6961 ccacgacaaa gattacagta atgttaatct gcatgagcat gccgaagcgc attctgagct
     7021 gccgaggcag gccaagagat ctatggcgga cgaggagaag ctgccgcccg gctgggagaa
     7081 gcgcatgagc cgcagctcag gccgagtgta ctacttcaac cacatcacta acgccagcca
     7141 gtgggagcgg cccagcggca acagcagcag tggtggcaaa aacgggcagg gggagcctgc
     7201 caggggtacc agtgctg
//
