LOCUS       sequence-208016-        6335 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vimentin fused to monomeric near-infrared fluorescent protein miRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6335)
  AUTHORS   Piatkevich KD, Jung EE, Straub C, Linghu C, Park D, Suk HJ, Hochbaum
            DR, Goodwin D, Pnevmatikakis E, Pak N, Kawashima T, Yang CT, Rhoades
            JL, Shemesh O, Asano S, Yoon YG, Freifeld L, Saulnier JL, Riegler C,
            Engert F, Hughes T, Drobizhev M, Szabo B, Ahrens MB, Flavell SW, 
            Sabatini BL, Boyden ES
  TITLE     A robotic multidimensional directed evolution approach applied to 
            fluorescent voltage reporters.
  JOURNAL   Nat Chem Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9.
  PUBMED    29483642
REFERENCE   2  (bases 1 to 6335)
  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 Chem 
            Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9."
COMMENT     Sequence Label: pmiRFP-Vimentin-17-C
FEATURES             Location/Qualifiers
     source          1..6335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      606..615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3123..3244
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3160..3179)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3214..3233
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3251..3706)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3388..3409)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3600..3619
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3733..3837
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3839..4196
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3839..4030
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3862..3882)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4047..4182
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4231..5025
                     /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             4231..5025
                     /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(4285..4304)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4895..4914
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5213..5232)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5257..5304
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5633..6221
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6122..6141
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtgccac catggtcgca ggacacgcaa gtggaagccc cgactttgga accgccgatc
      661 cgagcgactg tgagagagaa gaaatccatc tggccggatc catccagcca cacggaacac
      721 tgctggtggt ctccgagcct gaccatagaa tcattcaggc atctgccaac gccgctgaat
      781 tcctgaatct gggaagtgtg ctgggagtcc ccctggctga gatcgacggg gatctgctga
      841 tcaagattct gccccacctg gaccctaccg cagaaggaat gccagtggcc gtcaggtgcc
      901 gcattggaaa cccctccaca gagtacgatg gactgatgca tcgaccacct gaaggaggac
      961 tgatcattga gctggaacgg gctgggccac ccattgatct gagcggaaca ctggctcccg
     1021 cactggagcg aattcgaact gcaggatccc tgagggctct gtgcgacgat actgcactgc
     1081 tgttccagca gtgtaccggc tacgaccgag tgatggtcta tcggttcgat gaacaggggc
     1141 acggagaggt gtattcagaa atacatgtga ctggactgga gagctacttt ggcaacaggt
     1201 atcctagctc cctcgtgcca cagatggccc ggagactgta cgaaagacag agggtgcgcg
     1261 tcctggtgga tgtctcttat cagcctgtgc cactggagcc aagactgagt cccctgactg
     1321 gcagggacct ggatatgtca gggtgttttc tgcgctctat gagtcctact cacctgcagt
     1381 tcctgaagaa tatgggcgtg cgggcaacac tggtggtcag cctggtggtc ggaggcaaac
     1441 tgtgggggct ggtgatctgc caccattatc tgccacgctt catccatttt gagctgcgag
     1501 caatttgtga gctgctggcc gaagctatcg caacccgcat tacagccctg agatctggca
     1561 gcggtggagg cagcgcatcc ggcggaagcg gaagctccac caggtccgtg tcctcgtcct
     1621 cctaccgcag gatgttcggc ggcccgggca ccgcgagccg gccgagctcc agccggagct
     1681 acgtgactac gtccacccgc acctacagcc tgggcagcgc gctgcgcccc agcaccagcc
     1741 gcagcctcta cgcctcgtcc ccgggcggcg tgtatgccac gcgctcctct gccgtgcgcc
     1801 tgcggagcag cgtgcccggg gtgcggctcc tgcaggactc ggtggacttc tcgctggccg
     1861 acgccatcaa caccgagttc aagaacaccc gcaccaacga gaaggtggag ctgcaggagc
     1921 tgaatgaccg cttcgccaac tacatcgaca aggtgcgctt cctggagcag cagaataaga
     1981 tcctgctggc cgagctcgag cagctcaagg gccaaggcaa gtcgcgcctg ggggacctct
     2041 acgaggagga gatgcgggag ctgcgccggc aggtggacca gctaaccaac gacaaagccc
     2101 gcgtcgaggt ggagcgcgac aacctggccg aggacatcat gcgcctccgg gagaaattgc
     2161 aggaggagat gcttcagaga gaggaagccg aaaacaccct gcaatctttc agacaggatg
     2221 ttgacaatgc gtctctggca cgtcttgacc ttgaacgcaa agtggaatct ttgcaagaag
     2281 agattgcctt tttgaagaaa ctccacgaag aggaaatcca ggagctgcag gctcagattc
     2341 aggaacagca tgtccaaatc gatgtggatg tttccaagcc tgacctcacg gctgccctgc
     2401 gtgacgtacg tcagcaatat gaaagtgtgg ctgccaagaa cctgcaggag gcagaagaat
     2461 ggtacaaatc caagtttgct gacctctctg aggctgccaa ccggaacaat gacgccctgc
     2521 gccaggcaaa gcaggagtcc actgagtacc ggagacaggt gcagtccctc acctgtgaag
     2581 tggatgccct taaaggaacc aatgagtccc tggaacgcca gatgcgtgaa atggaagaga
     2641 actttgccgt tgaagctgct aactaccaag acactattgg ccgcctgcag gatgagattc
     2701 agaatatgaa ggaggaaatg gctcgtcacc ttcgtgaata ccaagacctg ctcaatgtta
     2761 agatggccct tgacattgag attgccacct acaggaagct gctggaaggc gaggagagca
     2821 ggatttctct gcctcttcca aacttttcct ccctgaacct gagggaaact aatctggatt
     2881 cactccctct ggttgatacc cactcaaaaa ggacacttct gattaagacg gttgaaacta
     2941 gagatggaca ggttatcaac gaaacttctc agcatcacga tgaccttgaa taaggatcca
     3001 ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3061 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3121 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3181 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3241 cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3301 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3361 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3421 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3481 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3541 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3601 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     3661 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     3721 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3781 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     3841 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3901 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3961 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4021 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4081 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4141 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     4201 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4261 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4321 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4381 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     4441 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4501 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4561 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4621 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4681 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4741 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4801 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4861 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4921 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4981 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5041 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5101 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5161 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     5221 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     5281 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     5341 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     5401 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     5461 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     5521 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5581 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5641 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5701 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5761 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5821 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5881 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5941 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6001 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6061 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6121 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6181 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6241 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6301 ctgattctgt ggataaccgt attaccgcca tgcat
//
