LOCUS       sequence-314090-        6477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nuclear Envelope, Excitation: 555, Emission: 584.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6477)
  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 6477)
  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-LaminB1-10
FEATURES             Location/Qualifiers
     source          1..6477
                     /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"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             553..1284
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    3205..3326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3242..3261)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3296..3315
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3333..3788)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3470..3491)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3682..3701
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3815..3919
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3921..4278
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3921..4112
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3944..3964)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4129..4264
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4313..5107
                     /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             4313..5107
                     /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(4367..4386)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4977..4996
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5295..5314)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5339..5386
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5715..6303
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6204..6223
                     /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 ccggtcgcca ccatggtgtc taagggcgaa gagctgatta aggagaacat gcacatgaag
      601 ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc
      661 aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc
      721 ttcgccttcg acatcctggc taccagcttc atgtacggca gcagaacctt catcaaccac
      781 acccagggca tccccgattt ctttaagcag tccttccctg agggcttcac atgggagaga
      841 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
      901 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg
      961 atgcagaaga aaacactcgg ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc
     1021 ggcctggaag gcagaaccga catggccctg aagctcgtgg gcgggggcca cctgatctgc
     1081 aacttcaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc
     1141 tactatgtgg accacagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag
     1201 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaactt
     1261 aatggcatgg acgagctgta caagtccgga ctcagatctc gagctcaagc ttcgatggcg
     1321 actgcgaccc ccgtgccgcc gcggatgggc agccgcgctg gcggccccac cacgccgctg
     1381 agccccacgc gcctgtcgcg gctccaggag aaggaggagc tgcgcgagct caatgaccgg
     1441 ctggcggtgt acatcgacaa ggtgcgcagc ctggagacgg agaacagcgc gctgcagctg
     1501 caggtgacgg agcgcgagga ggtgcgcggc cgtgagctca ccggcctcaa ggcgctctac
     1561 gagaccgagc tggccgacgc gcgacgcgcg ctcgacgaca cggcccgcga gcgcgccaag
     1621 ctgcagatcg agctgggcaa gtgcaaggcg gaacacgacc agctgctcct caactatgct
     1681 aagaaggaat ctgatcttaa tggcgcccag atcaagcttc gagaatatga agcagcactc
     1741 aattcgaaag atgcagctct tgctactgca cttggtgaca aaaaaagttt agagggagat
     1801 ttggaggatc tgaaggatca gattgcccag ttggaagcct ccttagctgc agccaaaaaa
     1861 cagttagcag atgaaacttt acttaaagta gatttggaga atcgttgtca gagccttact
     1921 gaggacttgg agtttcgcaa aagcatgtat gaagaggaga ttaacgagac cagaaggaag
     1981 catgaaacgc gcttggtaga ggtggattct gggcgtcaaa ttgagtatga gtacaagctg
     2041 gcgcaagccc ttcatgagat gagagagcaa catgatgccc aagtgaggct gtataaggag
     2101 gagctggagc agacttacca tgccaaactt gagaatgcca gactgtcatc agagatgaat
     2161 acttctactg tcaacagtgc cagggaagaa ctgatggaaa gccgcatgag aattgagagc
     2221 ctttcatccc agctttctaa tctacagaaa gagtctagag catgtttgga aaggattcaa
     2281 gaattagagg acttgcttgc taaagaaaaa gacaactctc gtcgcatgct gacagacaaa
     2341 gagagagaga tggcggaaat aagggatcaa atgcagcaac agctgaatga ctatgaacag
     2401 cttcttgatg taaagttagc cctggacatg gaaatcagtg cttacaggaa actcttagaa
     2461 ggcgaagaag agaggttgaa gctgtctcca agcccttctt cccgtgtgac agtatcccga
     2521 gcatcctcaa gtcgtagtgt acgtacaact agaggaaagc ggaagagggt tgatgtggaa
     2581 gaatcagagg cgagtagtag tgttagcatc tctcattccg cctcagccac tggaaatgtt
     2641 tgcatcgaag aaattgatgt tgatgggaaa tttatccgct tgaagaacac ttctgaacag
     2701 gatcaaccaa tgggaggctg ggagatgatc agaaaaattg gagacacatc agtcagttat
     2761 aaatatacct caagatatgt gctgaaggca ggccagactg ttacaatttg ggctgcaaac
     2821 gctggtgtca cagccagccc cccaactgac ctcatctgga agaaccagaa ctcgtggggc
     2881 actggcgaag atgtgaaggt tatattgaaa aattctcagg gagaggaggt tgctcaaaga
     2941 agtacagtct ttaaaacaac catacctgaa gaagaggagg aggaggaaga agcagctgga
     3001 gtggttgttg aggaagaact tttccaccag cagggaaccc caagagcatc caatagaagc
     3061 tgtgcaatta tgtagggatc caccggatct agataactga tcataatcag ccataccaca
     3121 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3181 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3241 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3301 ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa
     3361 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3421 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3481 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3541 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3601 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3661 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3721 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3781 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3841 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3901 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3961 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4021 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4081 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4141 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4201 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4261 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     4321 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4381 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4441 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4501 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4561 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4621 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4681 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4741 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4801 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4861 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4921 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4981 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5041 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5101 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5161 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5221 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     5281 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5341 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5401 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5461 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     5521 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     5581 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5641 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5701 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5761 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5821 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5881 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5941 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6001 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6061 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6121 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6181 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6241 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6301 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6361 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
     6421 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg
//
