LOCUS       sequence-250407-        4474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  F-Actin labeling with near-infrared fluorescent protein 
            miRFP670nano.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4474)
  AUTHORS   Oliinyk OS, Shemetov AA, Pletnev S, Shcherbakova DM, Verkhusha VV
  TITLE     Smallest near-infrared fluorescent protein evolved from 
            cyanobacteriochrome as versatile tag for spectral multiplexing.
  JOURNAL   Nat Commun. 2019 Jan 17;10(1):279. doi: 10.1038/s41467-018-08050-8.
  PUBMED    30655515
REFERENCE   2  (bases 1 to 4474)
  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: "2019-01-17"; volume: "10(1):279. doi"; pages: " 
            10.1038/s41467-018-08050-8"
COMMENT     Sequence Label: pLifeAct-miRFP670nano
FEATURES             Location/Qualifiers
     source          1..4474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        88..391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        392..595
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     546..566
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      624..633
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             630..680
                     /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"
     polyA_signal    1289..1410
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1326..1345)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1380..1399
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1417..1872)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1554..1575)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1766..1785
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1899..2003
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2005..2362
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2005..2196
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2028..2048)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2213..2348
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2397..3191
                     /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             2397..3191
                     /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(2451..2470)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3061..3080
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3379..3398)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3423..3470
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3799..4387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4288..4307
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
       61 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      121 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      181 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      241 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      301 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      361 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      421 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      481 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      541 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
      601 agtgaaccgt cagatccgct agcgccacca tgggcgtggc cgacttgatc aagaagttcg
      661 agtccatctc caaggaggag ggggatccac cggtaatggc aaacctggac aagatgctga
      721 ataccacagt aacagaggtg cggcagttcc tgcaggtgga cagagtgtgc gtgttccagt
      781 ttgaggagga ttatagcgga gtggtggtgg tggaggccgt ggacgatagg tggatctcca
      841 tcctgaagac ccaggtgcgg gatagatact tcatggagac aaggggcgag gagtattctc
      901 acggccgcta ccaggccatc gccgacatct acaccgcaaa cctgacagag tgctacaggg
      961 atctgctgac acagtttcag gtgagagcaa tcctggccgt gcccatcctg cagggcaaga
     1021 agctgtgggg cctgttggtg gcacaccagc tggcggcccc tagacagtgg cagacctggg
     1081 agatcgactt tctgaagcag caggccgtgg tggtgggcat cgccatccag cagagctagc
     1141 tgaattctgc agatatccag cacagtggcg gccgcgactc tagatcataa tcagccatac
     1201 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     1261 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     1321 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1381 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     1441 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1501 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1561 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1621 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1681 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1741 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     1801 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     1861 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1921 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1981 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     2041 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2101 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2161 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2221 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2281 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     2341 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     2401 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2461 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2521 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     2581 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2641 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     2701 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     2761 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     2821 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     2881 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     2941 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3001 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3061 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3121 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3181 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3241 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3301 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3361 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     3421 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     3481 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3541 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3601 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     3661 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3721 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3781 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3841 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3901 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3961 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4021 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4081 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4141 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4201 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4261 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4321 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4381 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4441 tcacatgttc tttcctgcgt tatcccctga ttct
//
