LOCUS       sequence-176879-        7526 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Negative-control mutant of FLINC-AKAR1-CAAX..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7526)
  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 7526)
  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: FLINC-AKAR1(TA)-CAAX
FEATURES             Location/Qualifiers
     source          1..7526
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             195..884
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTDMALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGH"
     promoter        complement(951..969)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(952..969)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(989..1006)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    995..1219
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1265..1693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1352..1371)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1562..1583
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1702..1722)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1707..2036
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1887..2022
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1949..1968
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2103..2897
                     /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(2157..2176)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2767..2786
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3071..3192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3108..3127)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3162..3181
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3241..3257)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3241..3257)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3254..3276)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3265..3281
                     /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(3289..3295,3296..3313,3314..3319))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3334..3355
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3472..3489)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3643..4231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3723..3742)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4402..5193,5194..5262))
                     /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     5025..5044
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5263..5367)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5442..5461)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5633..6012
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6013..6216
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6167..6187
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6261..6280
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6261..6279
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6299..6308
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6320..6337
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6341..6373
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(6377..6385,6386..6400)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             6407..7078
                     /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 ccggattgag gcgcgcggcc ctggttgacg
      181 gcggcagcga gctcatgagc gagctgatta aggagaacat gcacatgaag ctgtacatgg
      241 agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg
      301 agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg
      361 acatcctggc taccagcttc atgtacggca gcagaacctt catcaaccac acccagggca
      421 tccccgattt ctttaagcag tccttccctg agggcttcac atgggagaga gtcaccacat
      481 acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca
      541 tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg atgcagaaga
      601 aaacactcgg ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc ggcctggaag
      661 gcagaaccga catggccctg aagctcgtgg gcgggggcca cctgatctgc aacttcaaga
      721 ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc tactatgtgg
      781 accacagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag cagcacgagg
      841 tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaggaa ttcaagaaga
      901 agaagaagag caagaccaag tgcgtgatca tgtaatctag agggccctat tctatagtgt
      961 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     1021 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     1081 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     1141 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     1201 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     1261 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1321 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1381 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1441 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1501 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1561 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1621 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1681 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     1741 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     1801 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1861 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     1921 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     1981 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2041 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     2101 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2161 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2221 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2281 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2341 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2401 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2461 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2521 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2581 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     2641 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     2701 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     2761 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     2821 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     2881 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     2941 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3001 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3061 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3121 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3181 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     3241 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3301 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3361 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3421 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3481 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3541 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3601 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3661 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3721 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3781 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3841 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3901 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3961 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4021 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4081 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4141 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4201 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4261 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4321 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4381 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4441 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4501 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4561 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4621 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4681 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4741 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4801 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4861 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4921 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4981 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5041 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5101 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5161 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5221 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5281 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5341 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     5401 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     5461 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     5521 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     5581 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     5641 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     5701 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     5761 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     5821 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     5881 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     5941 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     6001 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     6061 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     6121 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     6181 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
     6241 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttgcggccgc
     6301 caccatggtg cggggttctc atcatcatca tcatcatggt atggctagca tgactggtgg
     6361 acagcaaatg ggtcgggatc tgtacgacga tgacgataag gatcccatga gtgtgattaa
     6421 accagacatg aagatcaagc tgcgtatgga aggcgctgta aatggacacc cgttcgcgat
     6481 tgaaggagtt ggccttggga agcctttcga gggaaaacag agtatggacc ttaaagtcaa
     6541 agaaggcgga cctctgcctt tcgcctatga catcttgaca actgtgttct gttacggcaa
     6601 cagggtattc gccaaatacc cagaaaatat agtagactat ttcaagcagt cgtttcctga
     6661 gggctactct tgggaacgaa gcatgaatta cgaagacggg ggcatttgta acgcgacaaa
     6721 cgacataacc ctggatggtg actgttatat ctatgaaatt cgatttgatg gtgtgaactt
     6781 tcctgccaat ggtccagtta tgcagaagag gactgtgaaa tgggagccat ccactgagaa
     6841 attgtatgtg cgtgatggag tgctgaaggg tgatgttaac atggctctgt cgcttgaagg
     6901 aggtggccat taccgatgtg acttcaaaac tacttataaa gctaagaagg ttgtccagtt
     6961 gccagactat cactttgtgg accaccacat tgagattaaa agccacgaca aagattacag
     7021 taatgttaat ctgcatgagc atgccgaagc gcattctgag ctgccgaggc aggccaagag
     7081 atctatgcat aagttttctc aagaacagat cggcgaaaac attgtgtgca gggtcatttg
     7141 taccacgggt caaattccca tccgagattt gtcagctgat atttcacaag tgcttaagga
     7201 aaaacgatcc ataaagaaag tttggacatt tggtagaaac ccagcctgtg actatcattt
     7261 aggaaacatt tcaagactgt caaataagca tttccaaata ctactaggag aagacggtaa
     7321 ccttttattg aatgacattt ccactaatgg gacctggtta aatgggcaaa aagtcgagaa
     7381 gaacagcaat cagttactgt cccaaggtga tgaaataacc gttggtgtag gcgtggaatc
     7441 agatatttta tctctggtca ttttcataaa cgacaaattt aagcagtgcc tggagcagaa
     7501 caaagttgat cgcggtacca gtgctg
//
