LOCUS       sequence-203803-        5283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Peroxisomes, Excitation: 559, Emission: 600.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5283)
  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 5283)
  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-PMP-N-10
FEATURES             Location/Qualifiers
     source          1..5283
                     /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      555..564
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1170..1179
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1176..1886
                     /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    2011..2132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2048..2067)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2102..2121
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2139..2594)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2276..2297)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2488..2507
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2621..2725
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2727..3084
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2727..2918
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2750..2770)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2935..3070
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3119..3913
                     /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             3119..3913
                     /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(3173..3192)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3783..3802
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4101..4120)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4145..4192
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4521..5109
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5010..5029
                     /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 ccggactcag atctgccacc atggcgccgg ccgcgtccag gctgcgggcc gaagccgggc
      601 tcggggcgct gccgcggcgg gcgctcgccc agtacttgct cttcctgcgg ctctacccgg
      661 tgctcaccaa ggcggccacc agtggcattt tgtcagcact tgggaacttc ctggcccaga
      721 tgattgagaa gaagcggaaa aaagaaaact ctagaagtct ggatgtcggt gggcctctga
      781 gatatgccgt ttacgggttc ttcttcacag ggccgctgag tcacttcttc tacttcttca
      841 tggaacattg gatccctcct gaggtccccc tggcagggct caggaggctt ctcctggacc
      901 gcctcgtctt tgcaccggcc ttcctcatgt tgttcttcct catcatgaac tttctggagg
      961 ggaaagacgc ctcagccttc gccgccaaga tgaggggggg cttctggccg gcgctgagga
     1021 tgaactggcg ggtgtggacg ccactacagt tcatcaacat caactacgtc cctctgaagt
     1081 tccgggtgct cttcgccaac ctggcagctc tgttctggta tgcctacctg gcctccttgg
     1141 ggaagagcgg ctccggtagc ggaccggtcg ccaccatggt gtctaagggc gaagagctga
     1201 tcaaggaaaa tatgcgtatg aaggtggtca tggaaggttc ggtcaacggc caccaattca
     1261 aatgcacagg tgaaggagaa ggcaatccgt acatgggaac tcaaaccatg aggatcaaag
     1321 tcatcgaggg aggacccctg ccatttgcct ttgacattct tgccacgtcg ttcatgtatg
     1381 gcagccgtac ttttatcaag tacccgaaag gcattcctga tttctttaaa cagtcctttc
     1441 ctgagggttt tacttgggaa agagttacga gatacgaaga tggtggagtc gtcaccgtca
     1501 tgcaggacac cagccttgag gatggctgtc tcgtttacca cgtccaagtc agaggggtaa
     1561 actttccctc caatggtccc gtgatgcaga agaagaccaa gggttgggag cctaatacag
     1621 agatgatgta tccagcagat ggtggtctga ggggatacac tcatatggca ctgaaagttg
     1681 atggtggtgg ccatctgtct tgctctttcg taacaactta caggtcaaaa aagaccgtcg
     1741 ggaacatcaa gatgcccggt atccatgccg ttgatcaccg cctggaaagg ttagaggaaa
     1801 gtgacaatga aatgttcgta gtacaacgcg aacacgcagt tgccaagttc gccgggcttg
     1861 gtggtgggat ggacgagctg tacaagtaag cggccgcgac tctagatcat aatcagccat
     1921 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     1981 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     2041 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     2101 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     2161 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     2221 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     2281 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     2341 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     2401 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     2461 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     2521 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     2581 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2641 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2701 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     2761 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2821 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2881 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2941 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3001 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     3061 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     3121 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3181 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3241 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3301 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3361 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3421 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3481 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3541 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3601 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     3661 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3721 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     3781 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3841 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     3901 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     3961 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4021 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4081 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     4141 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     4201 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     4261 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     4321 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     4381 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4441 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4501 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4561 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4621 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4681 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4741 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4801 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4861 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4921 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4981 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5041 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5101 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5161 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     5221 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     5281 ccg
//
