LOCUS       sequence-176183-        4707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Actin, Excitation: 588, Emission: 635.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4707)
  AUTHORS   Shcherbo D, Murphy CS, Ermakova GV, Solovieva EA, Chepurnykh TV, 
            Shcheglov AS, Verkhusha VV, Pletnev VZ, Hazelwood KL, Roche PM, 
            Lukyanov S, Zaraisky AG, Davidson MW, Chudakov DM
  TITLE     Far-red fluorescent tags for protein imaging in living tissues.
  JOURNAL   Biochem J. 2009 Mar 15;418(3):567-74. doi: 10.1042/BJ20081949.
  PUBMED    19143658
REFERENCE   2  (bases 1 to 4707)
  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: 
            "Biochem J"; date: "2009-03-15- 15"; volume: "418"; issue: "3"; 
            pages: "567-74"
COMMENT     Sequence Label: mKate1.31-Lifeact-7
FEATURES             Location/Qualifiers
     source          1..4707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     596..615
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(914..933)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    958..1005
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1334..1922
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1823..1842
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2097..2400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2401..2604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2555..2575
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2633..2642
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2639..2689
                     /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"
     regulatory      2705..2714
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2711..3409
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     polyA_signal    3531..3652
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3568..3587)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3622..3641
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3659..4114)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3796..3817)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4008..4027
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4141..4245
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4247..4604
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4247..4438
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4270..4290)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4455..4590
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(4639..4707,1..726)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             join(4639..4707,1..726)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(join(4693..4707,1..5))
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
       61 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
      121 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
      181 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
      241 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
      301 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
      361 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
      421 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
      481 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
      541 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
      601 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
      661 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
      721 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
      781 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
      841 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
      901 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
      961 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     1021 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     1081 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     1141 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     1201 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     1261 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1321 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1381 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1441 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1501 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1561 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1621 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1681 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1741 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1801 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1861 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1921 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1981 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     2041 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     2101 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     2161 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     2221 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     2281 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     2341 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     2401 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     2461 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     2521 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     2581 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgccaccat
     2641 gggcgtggcc gacttgatca agaagttcga gtccatctcc aaggaggagg gggatccacc
     2701 ggtcgccacc atggtgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga
     2761 gggcaccgtg aacaaccacc acttcaagtg cacatccgag ggcgaaggca agccctacga
     2821 gggcacccag accatgagaa tcaaggcggt cgagggcggc cctctcccct tcgccttcga
     2881 catcctggct accagcttca tgtacggcag caaaaccttc atcaaccaca cccagggcat
     2941 ccccgacttc tttaagcagt ccttccccga gggcttcaca tgggagagag tcaccacata
     3001 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     3061 ctacaacgtc aagatcagag gggtgaactt cccatccaac ggccctgtga tgcagaagaa
     3121 aacactcggc tgggaggcct ccaccgagac cctgtacccc gctgacggcg gcctggaagg
     3181 cagagccgac atggccctga agctcgtggg cgggggccac ctgatctgca acttgaagac
     3241 cacatacaga tccaagaaac ccgctaagaa cctcaagatg cccggcgtct actatgtgga
     3301 cagaagactg gaaagaatca aggaggccga caaagagacc tacgtcgagc agcacgaggt
     3361 ggctgtggcc agatactgcg acctccctag caaactgggg cacagatgag cggccgcgac
     3421 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3481 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3541 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3601 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     3661 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3721 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3781 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3841 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3901 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3961 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4021 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4081 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     4141 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4201 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     4261 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4321 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4381 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4441 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4501 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4561 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     4621 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4681 tgggtggaga ggctattcgg ctatgac
//
