LOCUS       sequence-270817-        4965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  F-actin labeling with a near-infrared fluorescent protein miRFP713.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4965)
  AUTHORS   Matlashov ME, Shcherbakova DM, Alvelid J, Baloban M, Pennacchietti 
            F, Shemetov AA, Testa I, Verkhusha VV
  TITLE     A set of monomeric near-infrared fluorescent proteins for multicolor
            imaging across scales.
  JOURNAL   Nat Commun. 2020 Jan 13;11(1):239. doi: 10.1038/s41467-019-13897-6.
  PUBMED    31932632
REFERENCE   2  (bases 1 to 4965)
  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"; journalName: "Nat 
            Commun."; date: "2020-01-13"; volume: "11(1):239. doi"; pages: " 
            10.1038/s41467-019-13897-6"
COMMENT     Sequence Label: pLifeAct-miRFP713
FEATURES             Location/Qualifiers
     source          1..4965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        83..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      619..628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             625..675
                     /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             706..1653
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="AEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGSD
                     NLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFIG
                     SWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKITG
                     FDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDIN
                     YRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGLI
                     VCHHRTPYYVDLDGRQACKRVAERLATQIGVMEE"
     polyA_signal    1775..1896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1812..1831)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1866..1885
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1903..2358)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2040..2061)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2252..2271
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2385..2489
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2491..2848
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2491..2682
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2514..2534)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2699..2834
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2883..3677
                     /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             2883..3677
                     /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(2937..2956)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3547..3566
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3865..3884)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3909..3956
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4285..4873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4774..4793
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      601 accgtcagat ccgctagcgc caccatgggc gtggccgact tgatcaagaa gttcgagtcc
      661 atctccaagg aggaggggga tccaccggtc ggtggcggtg gcagcgcgga aggctccgtc
      721 gccaggcagc ctgacctctt gacctgcgac gatgagccga tccatatccc cggtgccatc
      781 caaccgcatg gactgctgct cgccctcgcc gccgacatga cgatcgttgc cggcagcgac
      841 aaccttcccg aactcaccgg actggcgatc ggcgccctga tcggccgctc tgcggccgat
      901 gtcttcgact cggagacgca caaccgtctg acgatcgcct tggccgagcc cggggcggcc
      961 gtcggagcac cgatcactgt cggcttcacg atgcgaaagg acgcaggctt catcggctcc
     1021 tggcatcgcc atgatcagct catcttcctc gagctcgagc ctccccagcg ggacgtcgcc
     1081 gagccgcagg cgttcttccg ccgcaccaac agcgccatcc gccgcctgca ggccgccgaa
     1141 accttggaaa gcgcctgcgc cgccgcggcg caagaggtgc ggaagattac cggcttcgat
     1201 cgggtgatga tctatcgctt cgcctccgac ttcagcggcg aagtgatcgc agaggatcgg
     1261 tgcgccgagg tcgagtcaaa actaggcctg cactatcctg cctcaaccgt gccggcgcag
     1321 gcccgtcggc tctataccat caacccggta cggatcattc ccgatatcaa ttatcggccg
     1381 gtgccggtca ccccagacct caatccggtc accgggcggc cgattgatct tagcttcgcc
     1441 atcctgcgca gcgtctcgcc cgtccatctg gagttcatgc gcaacatagg catgcacggc
     1501 acgatgtcga tctcgatttt gcgcggcgag cgactgtggg gattgatcgt ttgccatcac
     1561 cgaacgccgt actacgtcga tctcgatggc cgccaagcct gcaagagggt cgccgagagg
     1621 ctggccactc agatcggcgt gatggaagag tgagcggccg cgactctaga tcataatcag
     1681 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     1741 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     1801 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     1861 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     1921 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1981 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2041 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2101 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     2161 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2221 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2281 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2341 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2401 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2461 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     2521 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2581 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     2641 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     2701 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     2761 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     2821 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     2881 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2941 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3001 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3061 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3121 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3181 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3241 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3301 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3361 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     3421 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3481 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     3541 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3601 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3661 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     3721 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3781 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3841 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     3901 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     3961 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4021 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4081 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     4141 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4201 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4261 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4321 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4381 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4441 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4501 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4561 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4621 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4681 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4741 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4801 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4861 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4921 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtgga
//
