LOCUS       sequence-271527-        6250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vimentin labeling with near-infrared fluorescent protein emiRFP703 
            (enhanced miRFP703).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6250)
  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 6250)
  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: pvimentin-emiRFP703
FEATURES             Location/Qualifiers
     source          1..6250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        114..417
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        418..621
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     572..592
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2103..2112
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3163..3284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3200..3219)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3254..3273
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3291..3746)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3428..3449)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3640..3659
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3773..3877
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3902..3922)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3946..4156
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3968..4164
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4015..4150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4199..4993
                     /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             4199..4993
                     /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(4253..4272)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4863..4882
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5181..5200)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5225..5272
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5601..6189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6090..6109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
       61 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      121 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      181 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      241 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      301 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      361 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      421 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      481 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      541 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      601 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
      661 cagatctcga gctcaagctt cgaattccca tgtccaccag gtccgtgtcc tcgtcctcct
      721 accgcaggat gttcggcggc ccgggcaccg cgagccggcc gagctccagc cggagctacg
      781 tgactacgtc cacccgcacc tacagcctgg gcagcgcgct gcgccccagc accagccgca
      841 gcctctacgc ctcgtccccg ggcggcgtgt atgccacgcg ctcctctgcc gtgcgcctgc
      901 ggagcagcgt gcccggggtg cggctcctgc aggactcggt ggacttctcg ctggccgacg
      961 ccatcaacac cgagttcaag aacacccgca ccaacgagaa ggtggagctg caggagctga
     1021 atgaccgctt cgccaactac atcgacaagg tgcgcttcct ggagcagcag aataagatcc
     1081 tgctggccga gctcgagcag ctcaagggcc aaggcaagtc gcgcctgggg gacctctacg
     1141 aggaggagat gcgggagctg cgccggcagg tggaccagct aaccaacgac aaagcccgcg
     1201 tcgaggtgga gcgcgacaac ctggccgagg acatcatgcg cctccgggag aaattgcagg
     1261 aggagatgct tcagagagag gaagccgaaa acaccctgca atctttcaga caggatgttg
     1321 acaatgcgtc tctggcacgt cttgaccttg aacgcaaagt ggaatctttg caagaagaga
     1381 ttgccttttt gaagaaactc cacgaagagg aaatccagga gctgcaggct cagattcagg
     1441 aacagcatgt ccaaatcgat gtggatgttt ccaagcctga cctcacggct gccctgcgtg
     1501 acgtacgtca gcaatatgaa agtgtggctg ccaagaacct gcaggaggca gaagaatggt
     1561 acaaatccaa gtttgctgac ctctctgagg ctgccaaccg gaacaatgac gccctgcgcc
     1621 aggcaaagca ggagtccact gagtaccgga gacaggtgca gtccctcacc tgtgaagtgg
     1681 atgcccttaa aggaaccaat gagtccctgg aacgccagat gcgtgaaatg gaagagaact
     1741 ttgccgttga agctgctaac taccaagaca ctattggccg cctgcaggat gagattcaga
     1801 atatgaagga ggaaatggct cgtcaccttc gtgaatacca agacctgctc aatgttaaga
     1861 tggcccttga cattgagatt gccacctaca ggaagctgct ggaaggcgag gagagcagga
     1921 tttctctgcc tcttccaaac ttttcctccc tgaacctgag ggaaactaat ctggattcac
     1981 tccctctggt tgatacccac tcaaaaagga cacttctgat taagacggtt gaaactagag
     2041 atggacaggt tatcaacgaa acttctcagc atcacgatga ccttgaaggg gatccaccgg
     2101 tcgccaccat ggcggaaggc tccgtcgcca ggcagcctga cctcttgacc tgcgaacatg
     2161 aagagatcca cctcgccggc tcgatccagc cgcatggcgc gcttctggtc gtcagcgaac
     2221 atgatcatcg cgtcatccag gccagcgcca acgccgcgga atttctgaat ctcggaagcg
     2281 tactcggcgt tccgctcgcc gagatcgacg gcgatctgtt gatcaagatc ctgccgcatc
     2341 tcgatcccac cgccgaaggc atgccggtcg cggtgcgctg ccggatcggc aatccctcta
     2401 cggagtactg cggtctgatg catcggcctc cggaaggcgg gctgatcatc gaactcgaac
     2461 gtgccggccc gtcgatcgat ctgtcaggca cgctggcgcc ggcgctggag cggatccgca
     2521 cggcgggttc actgcgcgcg ctgtgcgatg acaccgtgct gctgtttcag cagtgcaccg
     2581 gctacgaccg ggtgatggtg tatcgtttcg atgagcaagg ccacggcctg gtattctccg
     2641 agtgccatgt gcctgggctc gaatcctatt tcggcaaccg ctatccgtcg tcgctggtcc
     2701 cgcagatggc gcggcagctg tacgtgcggc agcgcgtccg cgtgctggtc gacgtcacct
     2761 atcagccggt gccgctggag ccgcggctgt cgccgctgac cgggcgcgat ctcgacatgt
     2821 cgggctgctt cctgcgctcg atgtcgccga tccatctgca gttcctgaag gacatgggcg
     2881 tgcgcgccac cctggcggtg tcgctggtgg tcggcggcaa gctgtggggc ctggttgtct
     2941 gtcaccatta tctgccgcgc ttcatccgtt tcgagctgcg ggcgatctgc aaacggctcg
     3001 ccgaaaggat cgcgacgcgg atcaccgcgc ttgagagcta agcggccgcg actctagatc
     3061 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3121 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3181 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3241 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     3301 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     3361 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     3421 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     3481 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     3541 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     3601 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     3661 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     3721 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     3781 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3841 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     3901 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3961 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4021 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4081 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4141 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4201 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4261 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4321 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4381 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4441 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4501 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4561 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4621 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4681 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4741 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4801 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4861 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4921 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4981 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5041 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5101 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5161 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5221 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5281 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5341 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5401 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5461 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5521 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5581 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5641 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5701 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5761 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5821 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5881 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5941 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6001 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6061 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6121 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6181 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6241 gctcacatgt
//
