LOCUS       sequence-221627-        5853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Mitochondria, Excitation: 559, Emission: 600.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5853)
  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 5853)
  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-PDHA1-N-10
FEATURES             Location/Qualifiers
     source          1..5853
                     /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      1740..1749
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1746..2456
                     /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    2581..2702
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2618..2637)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2672..2691
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2709..3164)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2846..2867)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3058..3077
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3191..3295
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3297..3654
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3297..3488
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3320..3340)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3505..3640
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3689..4483
                     /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             3689..4483
                     /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(3743..3762)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4353..4372
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4671..4690)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4715..4762
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5091..5679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5580..5599
                     /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 gctagcaccg
      541 ccaccatgag gaagatgctc gccgccgtct cccgcgtgct gtctggcgct tctcagaagc
      601 cggcaagcag agtgctggta gcatcccgta attttgcaaa tgatgctaca tttgaaatta
      661 agaaatgtga ccttcaccgg ctggaagaag gccctcctgt cacaacagtg ctcaccaggg
      721 aggatgggct caaatactac aggatgatgc agactgtacg ccgaatggag ttgaaagcag
      781 atcagctgta taaacagaaa attattcgtg gtttctgtca cttgtgtgat ggtcaggaag
      841 cttgctgtgt gggcctggag gccggcatca accccacaga ccatctcatc acagcctacc
      901 gggctcacgg ctttactttc acccggggcc tttccgtccg agaaattctc gcagagctta
      961 caggacgaaa aggaggttgt gctaaaggga aaggaggatc gatgcacatg tatgccaaga
     1021 acttctacgg gggcaatggc atcgtgggag cgcaggtgcc cctgggcgct gggattgctc
     1081 tagcctgtaa gtataatgga aaagatgagg tctgcctgac tttatatggc gatggtgctg
     1141 ctaaccaggg ccagatattc gaagcttaca acatggcagc tttgtggaaa ttaccttgta
     1201 ttttcatctg tgagaataat cgctatggaa tgggaacgtc tgttgagaga gcggcagcca
     1261 gcactgatta ctacaagaga ggcgatttca ttcctgggct gagagtggat ggaatggata
     1321 tcctgtgcgt ccgagaggca acaaggtttg ctgctgccta ttgtagatct gggaaggggc
     1381 ccatcctgat ggagctgcag acttaccgtt accacggaca cagtatgagt gaccctggag
     1441 tcagttaccg tacacgagaa gaaattcagg aagtaagaag taagagtgac cctattatgc
     1501 ttctcaagga caggatggtg aacagcaatc ttgccagtgt ggaagaacta aaggaaattg
     1561 atgtggaagt gaggaaggag attgaggatg ctgcccagtt tgccacggcc gatcctgagc
     1621 cacctttgga agagctgggc taccacatct actccagcga cccacctttt gaagttcgtg
     1681 gtgccaatca gtggatcaag tttaagtcag tcagtagctc cggaggggat ccaccggtcg
     1741 ccaccatggt gtctaagggc gaagagctga tcaaggaaaa tatgcgtatg aaggtggtca
     1801 tggaaggttc ggtcaacggc caccaattca aatgcacagg tgaaggagaa ggcaatccgt
     1861 acatgggaac tcaaaccatg aggatcaaag tcatcgaggg aggacccctg ccatttgcct
     1921 ttgacattct tgccacgtcg ttcatgtatg gcagccgtac ttttatcaag tacccgaaag
     1981 gcattcctga tttctttaaa cagtcctttc ctgagggttt tacttgggaa agagttacga
     2041 gatacgaaga tggtggagtc gtcaccgtca tgcaggacac cagccttgag gatggctgtc
     2101 tcgtttacca cgtccaagtc agaggggtaa actttccctc caatggtccc gtgatgcaga
     2161 agaagaccaa gggttgggag cctaatacag agatgatgta tccagcagat ggtggtctga
     2221 ggggatacac tcatatggca ctgaaagttg atggtggtgg ccatctgtct tgctctttcg
     2281 taacaactta caggtcaaaa aagaccgtcg ggaacatcaa gatgcccggt atccatgccg
     2341 ttgatcaccg cctggaaagg ttagaggaaa gtgacaatga aatgttcgta gtacaacgcg
     2401 aacacgcagt tgccaagttc gccgggcttg gtggtgggat ggacgagctg tacaagtaag
     2461 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     2521 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2581 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2641 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2701 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2761 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2821 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2881 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2941 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3001 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3061 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3121 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3181 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3241 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3301 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3361 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3421 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3481 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3541 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3601 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     3661 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3721 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3781 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3841 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     3901 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3961 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4021 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4081 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4141 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4201 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4261 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4321 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4381 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4441 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4501 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4561 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4621 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     4681 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4741 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4801 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4861 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4921 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     4981 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5041 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5101 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5161 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5221 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5281 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5341 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5401 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5461 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5521 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5581 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5641 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5701 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5761 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
     5821 gtcattagtt catagcccat atatggagtt ccg
//
