LOCUS       sequence-225631-        6107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  codes for a fusion of fluorescent protein mGarnet2 and lifeact that 
            targets F-actin in live cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6107)
  AUTHORS   Matela G, Gao P, Guigas G, Eckert AF, Nienhaus K, Nienhaus GU
  TITLE     A far-red emitting fluorescent marker protein, mGarnet2, for 
            microscopy and STED nanoscopy.
  JOURNAL   Chem Commun (Camb). 2017 Jan 10;53(5):979-982. doi: 
            10.1039/c6cc09081h.
  PUBMED    28044150
REFERENCE   2  (bases 1 to 6107)
  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"; journalName: 
            "Chem Commun (Camb)"; date: "2017-01-10- 10"; volume: "53"; issue: 
            "5"; pages: "979-982"
COMMENT     Sequence Label: LifeAct-mGarnet2
FEATURES             Location/Qualifiers
     source          1..6107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             667..717
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     CDS             724..1404
                     /codon_start=1
                     /product="monomeric variant of the red fluorescent protein 
                     eqFP611 (Kredel et al., 2009)"
                     /label=mRuby
                     /translation="MNSLIKENMRMKVVLEGSVNGHQFKCTGEGEGNPYMGTQTMRIKV
                     IEGGPLPFAFDILATSFMYGSKTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGVITV
                     MQDTSLEDGCLVYHAQLRGVNFPSNGAVMQKKTKGWEPNTEMMYPADGGLRGYNHMALK
                     VDGGGHLHCSLVTTYRSKKTVGNIKMPGIHAVDRRLERLEESDNEMFVVQREHAVAKFA
                     GLGGG"
     promoter        complement(1426..1444)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1427..1444)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1464..1481)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1470..1694
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1740..2168
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1827..1846)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2037..2058
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2177..2197)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2182..2511
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2362..2497
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2424..2443
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2578..3372
                     /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(2632..2651)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3242..3261
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3546..3667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3583..3602)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3637..3656
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3716..3732)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3716..3732)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3729..3751)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3740..3756
                     /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(3764..3770,3771..3788,3789..3794))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3809..3830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3947..3964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4118..4706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4198..4217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4877..5668,5669..5737))
                     /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     5500..5519
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5738..5842)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5917..5936)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccatgg gagtagcaga tctaatcaag aaattcgaga gcataagcaa agaggagctc
      721 gagatgaaca gcctgatcaa agaaaacatg cggatgaagg tggtgctgga aggcagcgtg
      781 aacggccacc agttcaagtg caccggcgag ggcgagggca acccctacat gggcacccag
      841 accatgcgga tcaaagtgat cgagggcgga cctctgccct tcgccttcga catcctggcc
      901 acatccttca tgtacggcag caagaccttc atcaagtacc ccaagggcat ccccgatttc
      961 ttcaagcaga gcttccccga gggcttcacc tgggagagag tgaccagata cgaggacggc
     1021 ggcgtgatca ccgtgatgca ggacaccagc ctggaagatg gctgcctggt gtaccatgcc
     1081 cagctcaggg gcgtgaattt tcccagcaac ggcgccgtga tgcagaagaa aaccaagggc
     1141 tgggagccca acaccgagat gatgtacccc gctgacggcg gactgagagg ctacaaccac
     1201 atggccctga aggtggacgg cggagggcac ctgcactgca gcctggtgac cacctaccgg
     1261 tccaagaaaa ccgtgggcaa catcaagatg cccggcatcc acgccgtgga ccggcggctg
     1321 gaaaggctgg aagagtccga caacgagatg ttcgtggtgc agcgggagca cgccgtggcc
     1381 aagttcgccg gcctgggcgg agggtgaaaa tctagagggc cctattctat agtgtcacct
     1441 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     1501 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     1561 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     1621 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     1681 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     1741 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1801 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1861 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1921 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1981 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2041 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2101 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2161 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2221 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2281 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     2341 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2401 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     2461 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     2521 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     2581 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     2641 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     2701 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     2761 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     2821 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     2881 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     2941 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3001 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3061 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     3121 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3181 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3241 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     3301 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     3361 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     3421 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     3481 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     3541 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3601 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3661 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     3721 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3781 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3841 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3901 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3961 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4021 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4081 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4141 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4201 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4261 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4321 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4381 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4441 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4501 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4561 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4621 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4681 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4741 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4801 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4861 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4921 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4981 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5041 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5101 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5161 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5221 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5281 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5341 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5401 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5461 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5521 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5581 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5641 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5701 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5761 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5821 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     5881 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     5941 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     6001 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     6061 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
