LOCUS       sequence-323958-        5745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry2-fused with a mitochondria-targeting signal of the
            ActA gene of Listeria monocytogenes under the control of CAG 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5745)
  AUTHORS   Tanida I, Furuta Y, Yamaguchi J, Kakuta S, Oliva Trejo JA, Uchiyama 
            Y
  TITLE     Two-color in-resin CLEM of Epon-embedded cells using osmium 
            resistant green and red fluorescent proteins.
  JOURNAL   Sci Rep. 2020 Dec 14;10(1):21871. doi: 10.1038/s41598-020-78879-x.
  PUBMED    33318540
REFERENCE   2  (bases 1 to 5745)
  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: "Sci Rep."; 
            date: "2020-12-14"; volume: "10(1):21871. doi"; pages: " 
            10.1038/s41598-020-78879-x"
COMMENT     Sequence Label: pCAG-mCherry2-mito
FEATURES             Location/Qualifiers
     source          1..5745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        53..328
                     /label=chicken beta-actin promoter
     intron          329..1345
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1269..1291
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1353..1372
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1409..1418
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1415..2122
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1560..1578)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2434..2453)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2499..2554
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2553..2572)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2915..2931)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2915..2931)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2928..2950)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2939..2955
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2963..2969,2970..2987,2988..2993))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3008..3029
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3088..3283
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3134..3269
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3289..3423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3326..3345)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3380..3399
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3491..3508)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3662..4250)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3742..3761)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4421..5212,5213..5281))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5044..5063
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5282..5386)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(5417..5745,1..51)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
       61 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
      121 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggcgcgcgcc
      181 aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc
      241 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
      301 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
      361 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
      421 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg
      481 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg
      541 cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
      601 ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg
      661 cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga
      721 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
      781 gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc
      841 ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg
      901 gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga
      961 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     1021 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga
     1081 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg
     1141 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct
     1201 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
     1261 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
     1321 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca
     1381 tcattttggc aaagctagcc taccggtcgc caccatggtg agcaagggcg aggaggataa
     1441 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     1501 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     1561 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     1621 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     1681 gctgtccttc cccgagggct tcaattggga gcgcgtgatg aacttcgagg acggcggcgt
     1741 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     1801 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag tgtcgtacca tgggctggga
     1861 ggcctccact gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     1921 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     1981 gaagcccgtg cagctgcccg gcgcctacaa cgtcgacatc aagttggaca tcctttccca
     2041 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     2101 cggcatggac gagctgtaca agtccggact cagatctaaa ctaattgcta aaagtgcaga
     2161 agacgaaaaa gcgaaggaag aaccagggaa ccatacgacg ttaattcttg caatgttagc
     2221 tattggcgtg ttctctttag gggcgtttat caaaattatt caattaagaa aaaataatta
     2281 agtcgagctc aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga
     2341 tctagataac tgatcaactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     2401 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     2461 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     2521 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     2581 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     2641 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     2701 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     2761 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     2821 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     2881 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2941 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     3001 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     3061 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
     3121 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3181 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3241 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
     3301 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3361 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     3421 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3481 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3541 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3601 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3661 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3721 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3781 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3841 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3901 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3961 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4021 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4081 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4141 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4201 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4261 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4321 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4381 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4441 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4501 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4561 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4621 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4681 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4741 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4801 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4861 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4921 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4981 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5041 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5101 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5161 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5221 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5281 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5341 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5401 aagtgccacc tgggtcgaca ttgattattg actagttatt aatagtaatc aattacgggg
     5461 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     5521 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     5581 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     5641 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     5701 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgg
//
