LOCUS       sequence-275041-        5936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Cytoskeleton, Excitation: 559, Emission: 600.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5936)
  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 5936)
  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-Myotilin-C-14
FEATURES             Location/Qualifiers
     source          1..5936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      25..613
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     514..533
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        899..1102
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1053..1073
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1141..1150
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1147..1857
                     /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    3523..3644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3560..3579)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3614..3633
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3651..4106)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3788..3809)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4000..4019
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4133..4237
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4262..4282)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4306..4516
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4328..4524
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4375..4510
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4559..5353
                     /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             4559..5353
                     /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(4613..4632)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5223..5242
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5541..5560)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5585..5632
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
       61 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
      121 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
      181 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
      241 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
      301 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
      361 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
      421 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
      481 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
      541 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
      601 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
      661 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
      721 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      781 agttccgcgt tacataactt acggtaaatg gcccgcctgg cattatgccc agtacatgac
      841 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      901 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      961 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     1021 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     1081 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc
     1141 gccaccatgg tgtctaaggg cgaagagctg atcaaggaaa atatgcgtat gaaggtggtc
     1201 atggaaggtt cggtcaacgg ccaccaattc aaatgcacag gtgaaggaga aggcaatccg
     1261 tacatgggaa ctcaaaccat gaggatcaaa gtcatcgagg gaggacccct gccatttgcc
     1321 tttgacattc ttgccacgtc gttcatgtat ggcagccgta cttttatcaa gtacccgaaa
     1381 ggcattcctg atttctttaa acagtccttt cctgagggtt ttacttggga aagagttacg
     1441 agatacgaag atggtggagt cgtcaccgtc atgcaggaca ccagccttga ggatggctgt
     1501 ctcgtttacc acgtccaagt cagaggggta aactttccct ccaatggtcc cgtgatgcag
     1561 aagaagacca agggttggga gcctaataca gagatgatgt atccagcaga tggtggtctg
     1621 aggggataca ctcatatggc actgaaagtt gatggtggtg gccatctgtc ttgctctttc
     1681 gtaacaactt acaggtcaaa aaagaccgtc gggaacatca agatgcccgg tatccatgcc
     1741 gttgatcacc gcctggaaag gttagaggaa agtgacaatg aaatgttcgt agtacaacgc
     1801 gaacacgcag ttgccaagtt cgccgggctt ggtggtggga tggacgagct gtacaagtcc
     1861 ggaagtgctg ggagcgcaag cggaagcgga ggctccggtt ttaactacga acgtccaaaa
     1921 cacttcatcc agtcccaaaa cccatgtggc tccagattgc agcctcctgg accagaaacc
     1981 tccagcttct ctagccagac caaacagtct tccattatca tccagccccg ccagtgtaca
     2041 gagcaaagat tttctgcctc ctcaacactg agctctcaca tcaccatgtc ctcctctgct
     2101 ttccctgctt ctccccagca gcatgctggc tccaacccag gccaaagggt tacaaccacc
     2161 tataaccagt ccccagccag cttcctcagc tccatattac catcacagcc tgattacaat
     2221 agcagtaaaa tcccttccgc tatggattcc aactatcaac agtcctcagc tggccaacct
     2281 ataaatgcaa agccatccca aactgcaaat gctaagccca taccaagaac tcctgatcat
     2341 gaaatacaag gatcaaaaga agctttgatt caagatttgg aaagaaagct gaaatgcaag
     2401 gacacccttc ttcataatgg aaatcaacgt ctaacatatg aagagaagat ggctcgcaga
     2461 ttgctaggac cacagaatgc agctgctgtg tttcaagctc aggatgacag tggtgcacaa
     2521 gactcgcagc aacacaactc agaacatgcg cgactgcaag ttcctacatc acaagtaaga
     2581 agtagatcaa cctcaagggg agatgtgaat gatcaggatg caatccagga gaaattttac
     2641 ccaccacgtt tcattcaagt gccagagaac atgtcgattg atgaaggaag attctgcaga
     2701 atggacttca aagtgagtgg actgccagct cctgatgtgt catggtatct aaatggaaga
     2761 acagttcaat cagatgattt gcacaaaatg atagtgtctg agaagggtct tcattcactc
     2821 atctttgaag tagtcagagc ttcagatgca ggggcttatg catgtgttgc caagaataga
     2881 gcaggagaag ccaccttcac tgtgcagctg gatgtccttg caaaagaaca taaaagagca
     2941 ccaatgttta tctacaaacc acagagcaaa aaagttttag agggagattc agtgaaacta
     3001 gaatgccaga tctcggctat acctccacca aagcttttct ggaaaagaaa taatgaaatg
     3061 gtacaattca acactgaccg aataagctta tatcaagata acactggaag agttacttta
     3121 ctgataaaag atgtaaacaa gaaagatgct gggtggtata ctgtgtcagc agttaatgaa
     3181 gctggagtga ctacatgtaa cacaagatta gacgttacgg cacgtccaaa ccaaactctt
     3241 ccagctccta agcagttacg ggttcgacca acattcagca aatatttagc acttaatggg
     3301 aaaggtttga atgtaaaaca agcttttaac ccagaaggag aatttcagcg tttggcagct
     3361 caatctggac tctatgaaag tgaagaactt taaggatcca ccggatctag ataactgatc
     3421 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3481 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3541 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3601 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     3661 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     3721 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     3781 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     3841 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     3901 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     3961 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     4021 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     4081 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     4141 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4201 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     4261 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4321 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4381 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4441 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4501 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4561 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4621 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4681 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4741 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4801 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4861 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4921 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4981 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5041 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     5101 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5161 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5221 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5281 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5341 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5401 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5461 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5521 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5581 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5641 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5701 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5761 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5821 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5881 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccc
//
