LOCUS       sequence-218671-        5450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Gap Junctions, Excitation: 555, Emission: 584.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5450)
  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 5450)
  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-Cx26-7
FEATURES             Location/Qualifiers
     source          1..5450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1434..1443
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1440..2171
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    2296..2417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2333..2352)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2387..2406
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2424..2879)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2561..2582)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2773..2792
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2906..3010
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3012..3369
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3012..3203
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3035..3055)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3220..3355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3404..4198
                     /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             3404..4198
                     /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(3458..3477)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4068..4087
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4386..4405)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4430..4477
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4806..5394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5295..5314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggactcagat ctcgagctca agcttcgaat tcgcggctgc acgctcctcg ggacacagtg
      721 ccaaccatcc agaggacaag atggattggg gcacactaca gagcatcctc gggggtgtca
      781 acaagcactc caccagcatt gggaaaatct ggctcactgt cctcttcatc ttccgcatca
      841 tgatcctcgt ggtggccgcg aaggaggtgt ggggagatga gcaagccgat tttgtttgca
      901 acactctcca gcctggctgt aagaatgtgt gctacgacca ctacttcccc atctctcaca
      961 tccggctctg ggctctgcag ctgatcatgg tgtccacgcc ggccctcctg gtagctatgc
     1021 acgtggccta ccggagacac gaaaagaaac ggaagttcat gaagggagag ataaagaacg
     1081 agtttaagga catcgaagag atcaaaaccc agaaggtccg tatcgaaggg tccctgtggt
     1141 ggacctacac caccagcatc ttcttccggg tcatcttcga agctgtcttc atgtatgtct
     1201 tttacatcat gtacaatggc ttcttcatgc agcgtctggt gaagtgtaac gcctggcctt
     1261 gtcccaatac agtggactgc ttcatttcca ggcccacaga aaagactgtc ttcacggtgt
     1321 tcatgatctc tgtgtctgga atttgcatcc tgctaaacat cacagagctg tgctatctgt
     1381 tcattaggta ttgctcaggg aagtccaaaa gaccagtcgc ggatccaccg gtcgccacca
     1441 tggtgtctaa gggcgaagag ctgattaagg agaacatgca catgaagctg tacatggagg
     1501 gcaccgtgaa caaccaccac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg
     1561 gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca
     1621 tcctggctac cagcttcatg tacggcagca gaaccttcat caaccacacc cagggcatcc
     1681 ccgatttctt taagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg
     1741 aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct
     1801 acaacgtcaa gatcagaggg gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa
     1861 cactcggctg ggaggccaac accgagatgc tgtaccccgc tgacggcggc ctggaaggca
     1921 gaaccgacat ggccctgaag ctcgtgggcg ggggccacct gatctgcaac ttcaagacca
     1981 catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcgtctac tatgtggacc
     2041 acagactgga aagaatcaag gaggccgaca aagagaccta cgtcgagcag cacgaggtgg
     2101 ctgtggccag atactgcgac ctccctagca aactggggca caaacttaat ggcatggacg
     2161 agctgtacaa gtaagcggcc gcgactctag atcataatca gccataccac atttgtagag
     2221 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     2281 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2341 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2401 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     2461 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     2521 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     2581 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     2641 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     2701 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     2761 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     2821 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     2881 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2941 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3001 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3061 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3121 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3181 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3241 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3301 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3361 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3421 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3481 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3541 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     3601 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3661 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     3721 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3781 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3841 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     3901 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     3961 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4021 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4081 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4141 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4201 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4261 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4321 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     4381 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     4441 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     4501 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     4561 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     4621 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     4681 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4741 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4801 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4861 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4921 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4981 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5041 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5101 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5161 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5221 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5281 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5341 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5401 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
//
