LOCUS       sequence-475401-        4932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Golgi Complex, Excitation: 555, Emission: 584.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4932)
  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 4932)
  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-Golgi-7
FEATURES             Location/Qualifiers
     source          1..4932
                     /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      798..807
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             804..1535
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    1660..1781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1697..1716)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1751..1770
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1788..2243)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1925..1946)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2137..2156
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2270..2374
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2376..2733
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2376..2567
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2399..2419)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2584..2719
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2768..3562
                     /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             2768..3562
                     /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(2822..2841)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3432..3451
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3750..3769)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3794..3841
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4170..4758
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4659..4678
                     /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 gctagcatga
      541 ggcttcggga gccgctcctg agcggcagcg ccgcgatgcc aggcgcgtcc ctacagcggg
      601 cctgccgcct gctcgtggcc gtctgcgctc tgcaccttgg cgtcaccctc gtttactacc
      661 tggctggccg cgacctgagc cgcctgcccc aactggtcgg agtctccaca ccgctgcagg
      721 gcggctcgaa cagtgccgcc gccatcgggc agtcctccgg ggagctccgg accggagggg
      781 ccaaggatcc accggtcgcc accatggtgt ctaagggcga agagctgatt aaggagaaca
      841 tgcacatgaa gctgtacatg gagggcaccg tgaacaacca ccacttcaag tgcacatccg
      901 agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg
      961 gccctctccc cttcgccttc gacatcctgg ctaccagctt catgtacggc agcagaacct
     1021 tcatcaacca cacccagggc atccccgatt tctttaagca gtccttccct gagggcttca
     1081 catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca
     1141 gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcccatcca
     1201 acggccctgt gatgcagaag aaaacactcg gctgggaggc caacaccgag atgctgtacc
     1261 ccgctgacgg cggcctggaa ggcagaaccg acatggccct gaagctcgtg ggcgggggcc
     1321 acctgatctg caacttcaag accacataca gatccaagaa acccgctaag aacctcaaga
     1381 tgcccggcgt ctactatgtg gaccacagac tggaaagaat caaggaggcc gacaaagaga
     1441 cctacgtcga gcagcacgag gtggctgtgg ccagatactg cgacctccct agcaaactgg
     1501 ggcacaaact taatggcatg gacgagctgt acaagtaagc ggccgcgact ctagatcata
     1561 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     1621 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     1681 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1741 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     1801 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     1861 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     1921 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     1981 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2041 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2101 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2161 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     2221 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2281 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2341 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     2401 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2461 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2521 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2581 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2641 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     2701 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     2761 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     2821 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     2881 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     2941 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3001 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3061 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3121 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3181 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3241 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     3301 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3361 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3421 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3481 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     3541 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     3601 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     3661 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     3721 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     3781 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     3841 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     3901 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     3961 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     4021 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4081 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4141 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4201 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4261 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4321 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4381 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4441 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4501 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4561 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4621 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4681 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4741 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4801 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4861 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     4921 tatggagttc cg
//
