LOCUS       sequence-247598-        9316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express FLN fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9316)
  TITLE     Knecht unpublished plasmids
REFERENCE   2  (bases 1 to 9316)
  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-mEos3.2
FEATURES             Location/Qualifiers
     source          1..9316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2581..3261
                     /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"
     CDS             complement(4066..4860)
                     /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(4177..4196)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4787..4806
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5556..5572)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5556..5572)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5569..5591)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5580..5596
                     /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(5604..5610,5611..5628,5629..5634))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5649..5670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5787..5804)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5958..6546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6038..6057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6717..7508,7509..7577))
                     /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     7340..7359
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7578..7682)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7750..7768
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7806..7828)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7928..7947
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atggctgctg ctccaagtgg aaaaacatgg attgatgtcc aaaaaaagac ttttacaggt
       61 tgggcaaata attatttaaa agaaagaatt ttaaagattg aagatcttgc aacaagttta
      121 gaagatggtg tacttttaat taatctttta gaaattattt caagtaaaaa gattttgaaa
      181 tataacaaag ctccaaagat tagaatgcaa aagattgaaa acaacaatat ggccgttaac
      241 tttatcaaga gtgaaggttt gaaattggtt ggtattggtg ctgaagatat cgttgatagt
      301 caattgaaat taattcttgg tttaatttgg accttgattt tacgttatca aattcaaatg
      361 tctgaaagtg ataactcacc aaaagcagct ctcttggaat gggttagaaa gcaagttgcc
      421 ccatacaagg ttgttgtaaa caatttcaca gactcatggt gtgatggtcg tgttctctct
      481 gctctcactg attccctcaa accaggtgtt agagagatgt caaccctcac cggtgatgcc
      541 gttcaagata tcgacagatc catggatatc gctcttgaag aatatgaaat cccaaagatt
      601 atggatgcca atgatatgaa ctctcttcca gatgaattat cagtcatcac ctacgtctct
      661 tatttccgtg attatgcact caacaaagaa aagagagatg ctgatgcttt agctgccctc
      721 gaaaagaaac gtcgtgaaac ttcagatgcc agcaaggttg aagtttatgg tccaggtgtt
      781 gaaggtggtt tcgttaataa atccgccgat ttccacatta aagccgtcaa ctactatggt
      841 gaaccattag ccaacggtgg tgaaggtttc actgttagcg tcgttggtgc cgatggtgtt
      901 gaagtcccat gcaaattggt tgacaacaaa aatggtatct atgatgcttc atacactgct
      961 accgttccac aagactacac tgtcgtcgtt caattagatg atgtccactg caaagactca
     1021 ccatacaacg ttaagattga cggttcagat gctcaacact caaatgccta cggtccaggt
     1081 ttagaaggtg gtaaagttgg tgttccagct gctttcaaaa tccaaggtcg taacaaggat
     1141 ggcgagaccg tcactcaagg tggtgatgat ttcaccgtca aagttcaatc accagaaggt
     1201 ccagtcgatg ctcaaatcaa agacaatggt gatggttcat acgatgttga atacaaacca
     1261 accaaaggtg gtgatcacac tgtcgaagtt ttcctccgtg gtgaaccatt agcccaaggt
     1321 ccaaccgaag ttaagatcct caactctgac tctcaaaaca gttattgtga tggtccaggt
     1381 ttcgaaaaag cccaagctaa acgtccaact gaattcacta ttcactctgt tggtgctgat
     1441 aataaaccat gtgctgctgg tggtgatcca ttccaagtat ccatcagtgg tccacaccca
     1501 gtcaacgttg gtatcaccga taatgacgat ggtacttaca ctgttgccta cactccagaa
     1561 caaccaggtg actacgaaat tcaagtcacc ctcaatgatg aagccatcaa ggatattcca
     1621 aaatcaattc atattaaacc agctgctgat ccagagaaat catacgctga aggtccaggt
     1681 ttagatggtg gtgaatgttt ccaaccaagc aaattcaaga ttcatgcagt tgatccagat
     1741 ggtgtacacc gtactgacgg tggtgacggt tttgtcgtta ccattgaagg tccagctcca
     1801 gtcgatccag ttatggtcga taatggtgat ggtacatatg atgttgaatt tgaaccaaaa
     1861 gaagccggtg actatgttat caatctcact ttagatggtg acaacgtcaa tggtttccca
     1921 aagactgtta ccgttaaacc agccccatcc gctgaacact cttatgctga aggtgaaggt
     1981 ttagtcaaag tatttgataa tgccccagct gaattcacta ttttcgccgt tgacactaaa
     2041 ggtgttgctc gtaccgatgg tggtgatcca tttgaagtcg ctatcaatgg tccagatggt
     2101 ttagtcgttg atgccaaagt taccgataac aatgacggta cttatggtgt tgtctatgat
     2161 gccccagttg aaggtaacta caatgttaat gtcaccctcc gtggtaatcc aatcaaaaat
     2221 atgccaatcg atgtcaaatg cattgaaggt gccaatggtg aagattcatc attcggttca
     2281 ttcactttta ccgtcgctgc taaaaataag aaaggtgaag ttaaaaccta tggtggtgat
     2341 aaatttgaag tctctatcac tggtccagct gaagaaatca ctctcgatgc tattgataac
     2401 caagatggta cttatactgc cgcttactct ttagttggta atggtcgttt ctcaactggt
     2461 gtcaaattaa acggtaaaca cattgaaggt tctccattca aacaagtact tggtaaccca
     2521 ggtaaaaaga atccagaagt taaatctttc actactactc gtactgccaa tctaggatcc
     2581 atgagtgcga ttaagccaga catgaagatc aaactccgta tggaaggcaa cgtaaacggg
     2641 caccactttg tgatcgacgg agatggtaca ggcaagcctt ttgagggaaa acagagtatg
     2701 gatcttgaag tcaaagaggg cggacctctg ccttttgcct ttgatatcct gaccactgca
     2761 ttccattacg gcaacagggt attcgccaaa tatccagaca acatacaaga ctattttaag
     2821 cagtcgtttc ctaaggggta ttcgtgggaa cgaagcttga ctttcgaaga cgggggcatt
     2881 tgcaacgcca gaaacgacat aacaatggaa ggggacactt tctataataa agttcgattt
     2941 tatggtacca actttcccgc caatggtcca gttatgcaga agaagacgct gaaatgggag
     3001 ccctccactg agaaaatgta tgtgcgtgat ggagtgctga cgggtgatgt agagatggct
     3061 ttgttgcttg aaggaaatgc ccattaccga tgtgacttca gaactactta caaagctaag
     3121 gagaagggtg tcaagttacc aggcgcccac tttgtggacc actgcattga gattttaagc
     3181 catgacaaag attacaacaa ggttaagctg tatgagcatg ctgttgctca ttctggattg
     3241 cctgacaatg ccagacgata actcgagtaa tctagaggat caattcaagt tttaccataa
     3301 aataaaaaaa aaaaataaaa aaaacaaatc caattatttt tatattaaat aattaaaata
     3361 aatgtaacaa taacaataaa tataataata tatacatata tatacaaaca ctatttcttt
     3421 tttttttttt ttttttagat ttaataaaaa attttattta tttttaaatt tttttttttt
     3481 tttttttttt tttttttttc taaactaaat atttggtata aatttaaatt atactagttg
     3541 aaacaaagtt tgatacacca catgtattct ttcctcttct taaataaatg tatccttgtt
     3601 ctccccaact tttttttttt tttaacaaaa ttaattatta gtaaactaaa ttattatttt
     3661 ttttattatt attattttaa aatttaaaat acttactctg caccccaaga attgatccgg
     3721 gaccaaagcg gccatcgtgc ctccccactc ctgcagttcg ggggcatgga tgcgcggata
     3781 gccgctgctg gtttcctgga tgccgacgga tttgcactgc cggtagaact ccgcgaggtc
     3841 gtccagcctc aggcagcagc tgaaccaact cgcgagggga tcgagcccgg ccggggtggg
     3901 cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa
     3961 cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca
     4021 ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc
     4081 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     4141 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     4201 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     4261 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
     4321 gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     4381 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     4441 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     4501 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     4561 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     4621 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg
     4681 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     4741 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     4801 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     4861 gcgaaacgat ccagcttgaa catcttcacc atccatttta tattatattt atttatttga
     4921 ttattttttg aattaattaa aaaaaaaaaa aaatttcatt ttataatctc agaaacctca
     4981 aaaaaaaaaa aataaaaaat aaaaaatata aaaaaataaa aataaaatcc caattttaaa
     5041 gcgaaaaacc acccatggtt tgaaaatttc aatcaatttc aaataacttt acttaaaaaa
     5101 aacccatttt ttatttaaaa atttcttttt taattaggca tcagaatatc aattctttgt
     5161 ttattacttg aattattttt ataattatta ttttttcata tctctttttt tttttatata
     5221 aaatatccat cattattctt caaaataaaa aaataattac aaaaagtgaa taataattaa
     5281 ataattaata aacctttaca cttttttttt tctttttaca gtttttatat ttattttaat
     5341 tttattattt tgcgcaactt tgaaaaaata aaaaagataa agtttttata aatatctatc
     5401 aaaaagatat cgttgttgat tgattatttt ggataatttt tctttaaatt ggggcaaatc
     5461 tgtaattttc agattaataa gatttttcct ttttattttt ctttttagtt tttgtaattt
     5521 ttaaaaataa acctctgaat taattcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5581 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5641 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5701 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5761 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5821 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5881 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5941 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6001 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6061 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6121 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6181 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6241 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6301 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6361 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6421 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6481 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6541 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6601 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6661 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6721 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6781 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6841 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6901 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6961 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7021 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7081 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7141 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7201 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7261 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7321 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7381 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7441 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7501 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7561 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7621 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7681 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     7741 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     7801 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     7861 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     7921 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     7981 cacagatgcg taaggagaaa ataccgcatc aggcgattcc aacatccaat aaatcataca
     8041 ggcaaggcaa agaattagca aaattaagca ataaagcctc agagcataaa gctaaatcgg
     8101 ttgtaccaaa aacattatga ccctgtaata cttttgcggg agaagccttt atttcaacgc
     8161 aaggataaaa atttttagaa ccctcatata ttttaaatgc aatgcctgag taatgtgtag
     8221 gtaaagattc aaacgggtga gaaaggccgg agacagtcaa atcaccatca atatgatatt
     8281 caaccgttct agctgataaa ttcatgccgg agagggtagc tatttttgag aggtctctac
     8341 aaaggctatc aggtcattgc ctgagagtct ggagcaaaca agagaatcga taggcaattt
     8401 atttttatat ctatctaaaa aaaaactagg aaaaatgaat gtcatcaaat agtattttaa
     8461 catttttttt ttttttttaa aaaaaaagtg taataaaaaa aaaaaaaaag tttaataaaa
     8521 aaaaaaaaaa aaaaaaagag gggaaagtaa ttataactag gttagttttt tataattttt
     8581 acatatttgt taataacttt taattttgaa tcatatgata ttacatcgtc ccgttgaaaa
     8641 aaaaaaaaaa aaattttttt tttcaaacat tttcattttt taaaaaatga tataaaattt
     8701 taaactaaac tattttatta aatacaaaca tataacttta tcttaatcaa tttttttggt
     8761 ttatacatat ttatgttcgt actgaagtat agatcttatt actaaagttt caaaagtttt
     8821 aaaaaaaatt aaagggggta aatatataac tttctgtttt tttcaattct gtcatgacaa
     8881 aaaaaaaaaa gtgtcatgac aaaaaaaaaa aaaaaaaaaa tttatttctt caataggtat
     8941 tgaaatgacc tccgttttta ataaaaagta tatatttgtg ctcgtctagg atctagctta
     9001 ctagtgtcga ctagataaaa aaaattttta tttattttta tttattttga attaaataga
     9061 ttacaaatta attaatccca tcaaatcttt aaaaaaaaaa atggtttaaa aaaacttggg
     9121 ttggttaatt attatttgaa aattttaaaa cccaaattaa aaaaaaaaaa tgggattcaa
     9181 aaattttttt tttttttttt tttttttttt tttttttttt tttttttttc ttatttctta
     9241 aaacaaataa attaaattaa ataaaaaata aaaatcagat cccaagctta aaccccgggc
     9301 tgcaggaatt ccgaaa
//
