LOCUS       sequence-492084-        6402 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  Localization: Intermediate Filaments, Excitation: 555, Emission: 
            584.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6402)
  AUTHORS   Shaner NC, Lin MZ, McKeown MR, Steinbach PA, Hazelwood KL, Davidson 
            MW, Tsien RY
  TITLE     Improving the photostability of bright monomeric orange and red 
            fluorescent proteins.
  JOURNAL   Nat Methods. 2008 Jun;5(6):545-51. doi: 10.1038/nmeth.1209. Epub 
            2008 May 4.
  PUBMED    18454154
REFERENCE   2  (bases 1 to 6402)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nmeth.1209"; journalName: "Nat Methods"; date: "2008-06"; 
            volume: "5"; issue: "6"; pages: "545-51"
COMMENT     Sequence Label: mTagRFP-T-Vimentin-7
FEATURES             Location/Qualifiers
     source          1..6402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    531..611
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      613..622
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        735..786
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     regulatory      2268..2277
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2274..3005
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    3130..3251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3167..3186)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3221..3240
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3258..3713)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3395..3416)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3607..3626
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3740..3844
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3846..4203
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3846..4037
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3869..3889)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4054..4189
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4238..5032
                     /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             4238..5032
                     /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(4292..4311)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4902..4921
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5220..5239)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5264..5311
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5640..6228
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6129..6148
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      601 gatccaccgg tcgccaccat ggtgtctaag ggcgaagagc tgattaagga gaacatgcac
      661 atgaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc
      721 gaaggcgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      781 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggactcagat ctcgagctca
      841 agcttcgaat tcccatgtcc accaggtccg tgtcctcgtc ctcctaccgc aggatgttcg
      901 gcggcccggg caccgcgagc cggccgagct ccagccggag ctacgtgact acgtccaccc
      961 gcacctacag cctgggcagc gcgctgcgcc ccagcaccag ccgcagcctc tacgcctcgt
     1021 ccccgggcgg cgtgtatgcc acgcgctcct ctgccgtgcg cctgcggagc agcgtgcccg
     1081 gggtgcggct cctgcaggac tcggtggact tctcgctggc cgacgccatc aacaccgagt
     1141 tcaagaacac ccgcaccaac gagaaggtgg agctgcagga gctgaatgac cgcttcgcca
     1201 actacatcga caaggtgcgc ttcctggagc agcagaataa gatcctgctg gccgagctcg
     1261 agcagctcaa gggccaaggc aagtcgcgcc tgggggacct ctacgaggag gagatgcggg
     1321 agctgcgccg gcaggtggac cagctaacca acgacaaagc ccgcgtcgag gtggagcgcg
     1381 acaacctggc cgaggacatc atgcgcctcc gggagaaatt gcaggaggag atgcttcaga
     1441 gagaggaagc cgaaaacacc ctgcaatctt tcagacagga tgttgacaat gcgtctctgg
     1501 cacgtcttga ccttgaacgc aaagtggaat ctttgcaaga agagattgcc tttttgaaga
     1561 aactccacga agaggaaatc caggagctgc aggctcagat tcaggaacag catgtccaaa
     1621 tcgatgtgga tgtttccaag cctgacctca cggctgccct gcgtgacgta cgtcagcaat
     1681 atgaaagtgt ggctgccaag aacctgcagg aggcagaaga atggtacaaa tccaagtttg
     1741 ctgacctctc tgaggctgcc aaccggaaca atgacgccct gcgccaggca aagcaggagt
     1801 ccactgagta ccggagacag gtgcagtccc tcacctgtga agtggatgcc cttaaaggaa
     1861 ccaatgagtc cctggaacgc cagatgcgtg aaatggaaga gaactttgcc gttgaagctg
     1921 ctaactacca agacactatt ggccgcctgc aggatgagat tcagaatatg aaggaggaaa
     1981 tggctcgtca ccttcgtgaa taccaagacc tgctcaatgt taagatggcc cttgacattg
     2041 agattgccac ctacaggaag ctgctggaag gcgaggagag caggatttct ctgcctcttc
     2101 caaacttttc ctccctgaac ctgagggaaa ctaatctgga ttcactccct ctggttgata
     2161 cccactcaaa aaggacactt ctgattaaga cggttgaaac tagagatgga caggttatca
     2221 acgaaacttc tcagcatcac gatgaccttg aaggggatcc accggtcgcc accatggtgt
     2281 ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
     2341 tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
     2401 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     2461 ctaccagctt catgtacggc agcagaacct tcatcaacca cacccagggc atccccgatt
     2521 tctttaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
     2581 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     2641 tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg
     2701 gctgggaggc caacaccgag atgctgtacc ccgctgacgg cggcctggaa ggcagaaccg
     2761 acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttcaag accacataca
     2821 gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg gaccacagac
     2881 tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag gtggctgtgg
     2941 ccagatactg cgacctccct agcaaactgg ggcacaaact taatggcatg gacgagctgt
     3001 acaagtaagc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     3061 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3121 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3181 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3241 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     3301 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3361 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3421 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3481 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3541 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3601 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3661 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     3721 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3781 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3841 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3901 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3961 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4021 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4081 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4141 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4201 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4261 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4321 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4381 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     4441 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4501 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4561 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4621 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4681 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4741 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     4801 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4861 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4921 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4981 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5041 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5101 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5161 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     5221 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     5281 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     5341 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     5401 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     5461 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     5521 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5581 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5641 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5701 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5761 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5821 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5881 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5941 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6001 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6061 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6121 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6181 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6241 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6301 ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc
     6361 aattacgggg tcattagttc atagcccata tatggagttc cg
//
