LOCUS       sequence-381313-        6075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Centrosomes, Excitation: 555, Emission: 584.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6075)
  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 6075)
  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"; doi: 
            "10.1038/nmeth.1209"; journalName: "Nat Methods"; date: "2008-06"; 
            volume: "5"; issue: "6"; pages: "545-51"
COMMENT     Sequence Label: mTagRFP-T-Gamma-Tubulin-17
FEATURES             Location/Qualifiers
     source          1..6075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             85..879
                     /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             85..879
                     /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(139..158)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     749..768
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1067..1086)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1111..1158
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1487..2075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1976..1995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2250..2553
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2554..2757
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2708..2728
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4190..4199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4196..4927
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    5052..5173
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5089..5108)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5143..5162
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5180..5635)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5317..5338)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5529..5548
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5662..5766
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5768..6075,1..50)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5768..5959
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5791..5811)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5976..6075,1..36)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
       61 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
      121 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
      181 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
      241 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
      301 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
      361 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
      421 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
      481 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
      541 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
      601 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
      661 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
      721 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
      781 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
      841 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
      901 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
      961 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     1021 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     1081 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     1141 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     1201 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     1261 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     1321 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1381 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1441 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1501 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1561 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1621 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1681 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1741 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1801 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1861 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1921 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1981 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2041 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2101 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2161 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     2221 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     2281 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     2341 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     2401 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     2461 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     2521 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     2581 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     2641 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     2701 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     2761 ttagtgaacc gtcagatccg ctagcgccac catgccgagg gaaatcatca ccctacagtt
     2821 gggccagtgc ggcaatcaga ttgggttcga gttctggaaa cagctgtgcg ccgagcatgg
     2881 tatcagcccc gagggcatcg tggaggagtt cgccaccgag ggcactgacc acaaggacgt
     2941 ctttttctac caggcagacg atgagcacta catcccccgg gccgtgctgc tggacttgga
     3001 accccgggtg atccactcca tcctcaactc cccctatgcc aagctctaca acccagagaa
     3061 catctacctg tcggaacatg gaggaggagc tggcaacaac tgggccagcg gattctccca
     3121 gggagaaaag atccatgagg acatttttga catcatagac cgggaggcag atggtagtga
     3181 cagtctagag ggctttgtgc tgtgtcactc cattgctggg gggacaggct ctggactggg
     3241 ttcctacctc ttagaacggc tgaatgacag gtatcctaag aagctggtgc agacatactc
     3301 agtgtttccc aaccaggacg agatgagcga tgtggtggtc cagccttaca attcactcct
     3361 cacactcaag aggctgacgc agaatgcaga ctgtgtggtg gtgctggaca acacagccct
     3421 gaaccggatt gccacagacc gcctgcacat ccagaaccca tccttctccc agatcaacca
     3481 gctggtgtct accatcatgt cagccagcac caccaccctg cgctaccctg gctacatgaa
     3541 caatgacctc atcggcctca tcgcctcgct cattcccacc ccacggctcc acttcctcat
     3601 gaccggctac acccctctca ctacggacca gtcagtggcc agcgtgagga agaccacggt
     3661 cctggatgtc atgaggcggc tgctgcagcc caagaacgtg atggtgtcca caggccgaga
     3721 ccgccagacc aaccactgct acatcgccat cctcaacatc atccagggag aggtggaccc
     3781 cacccaggtc cacaagagct tgcagaggat ccgggaacgc aagttggcca acttcatccc
     3841 gtggggcccc gccagcatcc aggtggccct gtcgaggaag tctccctacc tgccctcggc
     3901 ccaccgggtc agcgggctca tgatggccaa ccacaccagc atctcctcgc tcttcgagag
     3961 aacctgtcgc cagtatgaca agctgcgtaa gcgggaggcc ttcctggagc agttccgcaa
     4021 ggaggacatg ttcaaggaca actttgatga gatggacaca tccagggaga ttgtgcagca
     4081 gctcatcgat gagtaccatg cggccacacg gccagactac atctcctggg gcactcagga
     4141 gcagcgaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggtcg ccaccatggt
     4201 gtctaagggc gaagagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac
     4261 cgtgaacaac caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac
     4321 ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct
     4381 ggctaccagc ttcatgtacg gcagcagaac cttcatcaac cacacccagg gcatccccga
     4441 tttctttaag cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga
     4501 cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa
     4561 cgtcaagatc agaggggtga acttcccatc caacggccct gtgatgcaga agaaaacact
     4621 cggctgggag gccaacaccg agatgctgta ccccgctgac ggcggcctgg aaggcagaac
     4681 cgacatggcc ctgaagctcg tgggcggggg ccacctgatc tgcaacttca agaccacata
     4741 cagatccaag aaacccgcta agaacctcaa gatgcccggc gtctactatg tggaccacag
     4801 actggaaaga atcaaggagg ccgacaaaga gacctacgtc gagcagcacg aggtggctgt
     4861 ggccagatac tgcgacctcc ctagcaaact ggggcacaaa cttaatggca tggacgagct
     4921 gtacaagtaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     4981 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     5041 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5101 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5161 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     5221 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     5281 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     5341 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     5401 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     5461 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     5521 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     5581 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     5641 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5701 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5761 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5821 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5881 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5941 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     6001 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6061 cctcggcctc tgagc
//
