LOCUS       sequence-176124-        4722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Actin, Excitation: 559, Emission: 600.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4722)
  AUTHORS   Lam AJ, St-Pierre F, Gong Y, Marshall JD, Cranfill PJ, Baird MA, 
            McKeown MR, Wiedenmann J, Davidson MW, Schnitzer MJ, Tsien RY, Lin 
            MZ
  TITLE     Improving FRET dynamic range with bright green and red fluorescent 
            proteins.
  JOURNAL   Nat Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171.
  PUBMED    22961245
REFERENCE   2  (bases 1 to 4722)
  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"; journalName: "Nat 
            Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171."
COMMENT     Sequence Label: mRuby2-Lifeact-7
FEATURES             Location/Qualifiers
     source          1..4722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        56..247
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(79..99)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      264..399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             448..1242
                     /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             448..1242
                     /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(502..521)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1112..1131
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1430..1449)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1474..1521
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1850..2438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2339..2358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2613..2916
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2917..3120
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3071..3091
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3149..3158
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3155..3205
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     regulatory      3221..3230
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3227..3937
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     polyA_signal    4062..4183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4099..4118)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4153..4172
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4190..4645)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4327..4348)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4539..4558
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(4672..4722,1..54)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
       61 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
      121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
      181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      241 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
      301 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
      361 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
      421 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
      481 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
      541 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
      601 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
      661 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
      721 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
      781 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
      841 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
      901 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
      961 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     1021 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     1081 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     1141 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     1201 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     1261 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     1321 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     1381 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     1441 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     1501 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     1561 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     1621 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     1681 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1741 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1801 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1861 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1921 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1981 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2041 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2101 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2161 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2221 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2281 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2341 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2401 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2461 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2521 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
     2581 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     2641 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     2701 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     2761 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     2821 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     2881 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     2941 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     3001 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     3061 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     3121 ggtttagtga accgtcagat ccgctagcgc caccatgggc gtggccgact tgatcaagaa
     3181 gttcgagtcc atctccaagg aggaggggga tccaccggtc gccaccatgg tgtctaaggg
     3241 cgaagagctg atcaaggaaa atatgcgtat gaaggtggtc atggaaggtt cggtcaacgg
     3301 ccaccaattc aaatgcacag gtgaaggaga aggcaatccg tacatgggaa ctcaaaccat
     3361 gaggatcaaa gtcatcgagg gaggacccct gccatttgcc tttgacattc ttgccacgtc
     3421 gttcatgtat ggcagccgta cttttatcaa gtacccgaaa ggcattcctg atttctttaa
     3481 acagtccttt cctgagggtt ttacttggga aagagttacg agatacgaag atggtggagt
     3541 cgtcaccgtc atgcaggaca ccagccttga ggatggctgt ctcgtttacc acgtccaagt
     3601 cagaggggta aactttccct ccaatggtcc cgtgatgcag aagaagacca agggttggga
     3661 gcctaataca gagatgatgt atccagcaga tggtggtctg aggggataca ctcatatggc
     3721 actgaaagtt gatggtggtg gccatctgtc ttgctctttc gtaacaactt acaggtcaaa
     3781 aaagaccgtc gggaacatca agatgcccgg tatccatgcc gttgatcacc gcctggaaag
     3841 gttagaggaa agtgacaatg aaatgttcgt agtacaacgc gaacacgcag ttgccaagtt
     3901 cgccgggctt ggtggtggga tggacgagct gtacaagtaa gcggccgcga ctctagatca
     3961 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     4021 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     4081 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4141 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     4201 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4261 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4321 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4381 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4441 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4501 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4561 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4621 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4681 cctatttgtt tatttttcta aatacattca aatatgtatc cg
//
