LOCUS       sequence-211958-        4750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Mitochondria, Excitation: 587, Emission: 610.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4750)
  AUTHORS   Olenych SG, Claxton NS, Ottenberg GK, Davidson MW
  TITLE     The fluorescent protein color palette.
  JOURNAL   Curr Protoc Cell Biol. 2007 Sep;Chapter 21:Unit 21.5. doi: 
            10.1002/0471143030.cb2105s36.
  PUBMED    18228502
REFERENCE   2  (bases 1 to 4750)
  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: "Curr Protoc
            Cell Biol."; date: "2007-09"; volume: "Chapter 21:Unit 21.5. doi"; 
            pages: " 10.1002/0471143030.cb2105s36"
COMMENT     Sequence Label: mCherry-Mito-7
FEATURES             Location/Qualifiers
     source          1..4750
                     /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"
     CDS             537..611
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      639..648
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             645..1355
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(790..808)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1058..1077
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1478..1599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1515..1534)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1569..1588
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1606..2061)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1743..1764)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1955..1974
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2088..2192
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2194..2551
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2194..2385
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2217..2237)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2402..2537
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2586..3380
                     /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             2586..3380
                     /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(2640..2659)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3250..3269
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3568..3587)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3612..3659
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3988..4576
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4477..4496
                     /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 gctagcatgt
      541 ccgtcctgac gccgctgctg ctgcggggct tgacaggctc ggcccggcgg ctcccagtgc
      601 cgcgcgccaa gatccattcg ttgggggatc caccggtcgc caccatggtg agcaagggcg
      661 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
      721 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
      781 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
      841 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
      901 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
      961 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     1021 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     1081 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     1141 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     1201 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     1261 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     1321 actccaccgg cggcatggac gagctgtaca agtaaagcgg ccgcgactct agatcataat
     1381 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     1441 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1501 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1561 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     1621 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1681 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     1741 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     1801 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     1861 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     1921 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     1981 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     2041 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2101 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2161 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     2221 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2281 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2341 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2401 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2461 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     2521 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     2581 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     2641 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     2701 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     2761 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     2821 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     2881 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     2941 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3001 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3061 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     3121 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3181 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3241 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3301 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3361 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     3421 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     3481 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     3541 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     3601 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     3661 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     3721 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     3781 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     3841 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3901 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3961 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4021 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4081 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4141 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4201 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4261 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4321 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4381 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4441 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4501 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4561 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4621 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4681 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     4741 tggagttccg
//
