LOCUS       sequence-247612-        8650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express filamin with actin binding domain deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8650)
  TITLE     Knecht unpublished plasmids
REFERENCE   2  (bases 1 to 8650)
  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"
COMMENT     Sequence Label: FLN-deltaABD-mEos3.2
FEATURES             Location/Qualifiers
     source          1..8650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(594..1388)
                     /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(705..724)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1315..1334
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(2155..2171)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2155..2171)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2168..2190)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2179..2195
                     /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(2203..2209,2210..2227,2228..2233))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2248..2269
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2386..2403)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2557..3145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2637..2656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3316..4107,4108..4176))
                     /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     3939..3958
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4177..4281)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4349..4367
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4405..4427)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4527..4546
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             5971..5991
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             7831..8511
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.1
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDVEMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
ORIGIN
        1 tttttttttt tttttttttt tttttttcta aactaaatat ttggtataaa tttaaattat
       61 actagttgaa acaaagtttg atacaccaca tgtattcttt cctcttctta aataaatgta
      121 tccttgttct ccccaacttt tttttttttt taacaaaatt aattattagt aaactaaatt
      181 attatttttt ttattattat tattttaaaa tttaaaatac ttactctgca ccccaagaat
      241 tgatccggga ccaaagcggc catcgtgcct ccccactcct gcagttcggg ggcatggatg
      301 cgcggatagc cgctgctggt ttcctggatg ccgacggatt tgcactgccg gtagaactcc
      361 gcgaggtcgt ccagcctcag gcagcagctg aaccaactcg cgaggggatc gagcccggcc
      421 ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc
      481 ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg
      541 tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag
      601 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa
      661 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc
      721 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa
      781 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga
      841 tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc
      901 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct
      961 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc
     1021 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac
     1081 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca
     1141 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc
     1201 tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc
     1261 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag
     1321 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt
     1381 tcaatcatgc gaaacgatcc agcttgaaca tcttcaccat ccattttata ttatatttat
     1441 ttatttgatt attttttgaa ttaattaaaa aaaaaaaaaa atttcatttt ataatctcag
     1501 aaacctcaaa aaaaaaaaaa taaaaaataa aaaatataaa aaaataaaaa taaaatccca
     1561 attttaaagc gaaaaaccac ccatggtttg aaaatttcaa tcaatttcaa ataactttac
     1621 ttaaaaaaaa cccatttttt atttaaaaat ttctttttta attaggcatc agaatatcaa
     1681 ttctttgttt attacttgaa ttatttagga aaaaaaaaaa aaaaaaaaaa tcatccattt
     1741 gtactaaaac tttaaaattc tatttttttt tttttttttt ataattatta ttttttcata
     1801 tctctttttt ttttatataa aatatccatc attattcttc aaaataaaaa aataattaca
     1861 aaaagtgaat aataattaaa taattaataa acctttacac tttttttttt ctttttacag
     1921 tttttatatt tattttaatt ttattatttt gcgcaacttt gaaaaaataa aaaagataaa
     1981 gtttttataa atatctatca aaaagatatc gttgttgatt gattattttg gataattttt
     2041 ctttaaattg gggcaaatct gtaattttca gattaataag atttttcctt tttatttttc
     2101 tttttagttt ttgtaatttt taaaaataaa cctctgaatt aattcgtaat catggtcata
     2161 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2221 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2281 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2341 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2401 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2461 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2521 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2581 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2641 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2701 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2761 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2821 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2881 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2941 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3001 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3061 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3121 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3181 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3241 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3301 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3361 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3421 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3481 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3541 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3601 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3661 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3721 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3781 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3841 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3901 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3961 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4021 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4081 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4141 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4201 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4261 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4321 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     4381 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4441 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4501 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4561 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgattcca
     4621 acatccaata aatcatacag gcaaggcaaa gaattagcaa aattaagcaa taaagcctca
     4681 gagcataaag ctaaatcggt tgtaccaaaa acattatgac cctgtaatac ttttgcggga
     4741 gaagccttta tttcaacgca aggataaaaa tttttagaac cctcatatat tttaaatgca
     4801 atgcctgagt aatgtgtagg taaagattca aacgggtgag aaaggccgga gacagtcaaa
     4861 tcaccatcaa tatgatattc aaccgttcta gctgataaat tcatgccgga gagggtagct
     4921 atttttgaga ggtctctaca aaggctatca ggtcattgcc tgagagtctg gagcaaacaa
     4981 gagaatcgat aggcaattta tttttatatc tatctaaaaa aaaactagga aaaatgaatg
     5041 tcatcaaata gtattttaac attttttttt tttttttaaa aaaaaawktt watnnacaaa
     5101 aaaaaaaaaa wtttntatta naaaaaaaaa aaaaaaaaaa aaaaaaaaaa aakttaawaa
     5161 aaaaaaaaaa aaaaaaaaag aggggaaagt aattataact aggttagttt tttataattt
     5221 ttacatattt gttaataact tttaattttg aatcatatga tattacatcg tcccgttgaa
     5281 aaaaaaaaaa aaaaattttt tttttcaaac attttcattt tttaaaaaat gatataaaat
     5341 tttaaactaa actattttat taaatacaaa catataactt tatcttaatc aatttttttg
     5401 gtttatacat atttatgttc gtactgaagt atagatctta ttactaaagt ttcaaaagtt
     5461 ttaaaaaaaa ttaaaggggg taaatatata actttctgtt tttttcaatt ctgtcatgac
     5521 aaaaaaaaaa aagtgtcatg acaaaaaaaa aaaaaaaaaa atttatttct tcaataggta
     5581 ttgaaatgac ctccgttttt aataaaaagt atatatttgt gctcgtctag gatctagctt
     5641 actagtgtcg actagataaa aaaaattttt atttattttt atttattttg aattaaatag
     5701 attacaaatt aattaatccc atcaaatctt taaaaaaaaa aatggtttaa aaaaacttgg
     5761 gttggttaat tattatttga aaattttaaa acccaaatta aaaaaaaaaa atgggattca
     5821 aaaatttttt tttttttttt tttttttttt tttttttttt tttttttttt twttttataa
     5881 aaaaattttt tttttttttt ttcttatttc ttaaaacaaa taaattaaat taaataaaaa
     5941 ataaaaatca gatcccaagc ttaaaaaatg catcatcatc atcatcatca cgatggtacc
     6001 gagctcatgg ttgaagttta tggtccaggt gttgaaggtg gtttcgttaa taaatccgcc
     6061 gatttccaca ttaaagccgt caactactat ggtgaaccat tagccaacgg tggtgaaggt
     6121 ttcactgtta gcgtcgttgg tgccgatggt gttgaagtcc catgcaaatt ggttgacaac
     6181 aaaaatggta tctatgatgc ttcatacact gctaccgttc cacaagacta cactgtcgtc
     6241 gttcaattag atgatgtcca ctgcaaagac tcaccataca acgttaagat tgacggttca
     6301 gatgctcaac actcaaatgc ctacggtcca ggtttagaag gtggtaaagt tggtgttcca
     6361 gctgctttca aaatccaagg tcgtaacaag gatggcgaga ccgtcactca aggtggtgat
     6421 gatttcaccg tcaaagttca atcaccagaa ggtccagtcg atgctcaaat caaagacaat
     6481 ggtgatggtt catacgatgt tgaatacaaa ccaaccaaag gtggtgatca cactgtcgaa
     6541 gttttcctcc gtggtgaacc attagcccaa ggtccaaccg aagttaagat cctcaactct
     6601 gactctcaaa acagttattg tgatggtcca ggtttcgaaa aagcccaagc taaacgtcca
     6661 actgaattca ctattcactc tgttggtgct gataataaac catgtgctgc tggtggtgat
     6721 ccattccaag tatccatcag tggtccacac ccagtcaacg ttggtatcac cgataatgac
     6781 gatggtactt acactgttgc ctacactcca gaacaaccag gtgactacga aattcaagtc
     6841 accctcaatg atgaagccat caaggatatt ccaaaatcaa ttcatattaa accagctgct
     6901 gatccagaga aatcatacgc tgaaggtcca ggtttagatg gtggtgaatg tttccaacca
     6961 agcaaattca agattcatgc agttgatcca gatggtgtac accgtactga cggtggtgac
     7021 ggttttgtcg ttaccattga aggtccagct ccagtcgatc cagttatggt cgataatggt
     7081 gatggtacat atgatgttga atttgaacca aaagaagccg gtgactatgt tatcaatctc
     7141 actttagatg gtgacaacgt caatggtttc ccaaagactg ttaccgttaa accagcccca
     7201 tccgctgaac actcttatgc tgaaggtgaa ggtttagtca aagtatttga taatgcccca
     7261 gctgaattca ctattttcgc cgttgacact aaaggtgttg ctcgtaccga tggtggtgat
     7321 ccatttgaag tcgctatcaa tggtccagat ggtttagtcg ttgatgccaa agttaccgat
     7381 aacaatgacg gtacttatgg tgttgtctat gatgccccag ttgaaggtaa ctacaatgtt
     7441 aatgtcaccc tccgtggtaa tccaatcaaa aatatgccaa tcgatgtcaa atgcattgaa
     7501 ggtgccaatg gtgaagattc atcattcggt tcattcactt ttaccgtcgc tgctaaaaat
     7561 aagaaaggtg aagttaaaac ctatggtggt gataaatttg aagtctctat cactggtcca
     7621 gctgaagaaa tcactctcga tgctattgat aaccaagatg gtacttatac tgccgcttac
     7681 tctttagttg gtaatggtcg tttctcaact ggtgtcaaat taaacggtaa acacattgaa
     7741 ggttctccat tcaaacaagt acttggtaac ccaggtaaaa agaatccaga agttaaatct
     7801 ttcactacta ctcgtactgc caatggatcc atgagtgcga ttaagccaga catgaagatc
     7861 aaactccgta tggaaggcaa cgtaaacggg caccactttg tgatcgacgg agatggtaca
     7921 ggcaagcctt ttgagggaaa acagagtatg gatcttgaag tcaaagaggg cggacctctg
     7981 ccttttgcct ttgatatcct gaccactgca ttccattacg gcaacagggt attcgccaaa
     8041 tatccagaca acatacaaga ctattttaag cagtcgtttc ctaaggggta ttcgtgggaa
     8101 cgaagcttga ctttcgaaga cgggggcatt tgcaacgcca gaaacgacat aacaatggaa
     8161 ggggacactt tctataataa agttcgattt tatggtacca actttcccgc caatggtcca
     8221 gttatgcaga agaagacgct gaaatgggag ccctccactg agaaaatgta tgtgcgtgat
     8281 ggagtgctga cgggtgatgt agagatggct ttgttgcttg aaggaaatgc ccattaccga
     8341 tgtgacttca gaactactta caaagctaag gagaagggtg tcaagttacc aggcgcccac
     8401 tttgtggacc actgcattga gattttaagc catgacaaag attacaacaa ggttaagctg
     8461 tatgagcatg ctgttgctca ttctggattg cctgacaatg ccagacgata atctagagga
     8521 tcaattcaag ttttaccata aaataaaaaa aaaaaataaa aaaaacaaat ccaattattt
     8581 ttatattaaa taattaaaat aaatgtaaca ataacaataa atataataat atatacatat
     8641 atatacaaac
//
