LOCUS       sequence-323956-        5563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry2 under the control of CAG promoter, basic vector, 
            multiclonig site at C terminal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5563)
  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 5563)
  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-G
FEATURES             Location/Qualifiers
     source          1..5563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          152..1168
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1092..1114
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1176..1195
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1232..1241
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1238..1945
                     /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(1383..1401)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    2003..2059
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(2159..2178)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2224..2279
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2278..2297)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2640..2656)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2640..2656)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2653..2675)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2664..2680
                     /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(2688..2694,2695..2712,2713..2718))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2733..2754
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2813..3008
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2859..2994
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3014..3148
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3051..3070)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3105..3124
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3216..3233)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3387..3975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3467..3486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4146..4937,4938..5006))
                     /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     4769..4788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5007..5111)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5142..5521
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc
       61 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
      121 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt
      181 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc
      241 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg
      301 acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg agggcccttt
      361 gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
      421 cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct
      481 ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg
      541 cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg
      601 cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg agcacggccc
      661 ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc cgggcggggg
      721 gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg agggctcggg
      781 ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca
      841 ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc aaatctgtgc
      901 ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg cgaagcggtg
      961 cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc
     1021 ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc gggggggacg
     1081 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca
     1141 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc
     1201 tcatcatttt ggcaaagcta gcctaccggt cgccaccatg gtgagcaagg gcgaggagga
     1261 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     1321 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     1381 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     1441 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     1501 gaagctgtcc ttccccgagg gcttcaattg ggagcgcgtg atgaacttcg aggacggcgg
     1561 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     1621 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagtgtcgta ccatgggctg
     1681 ggaggcctcc actgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     1741 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     1801 caagaagccc gtgcagctgc ccggcgccta caacgtcgac atcaagttgg acatcctttc
     1861 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     1921 cggcggcatg gacgagctgt acaagtccgg actcaccggt ggaggcggtt caggcggagg
     1981 tggctctggc ggtggcggat cgagatctcg agctcaagct tcgaattctg cagtcgacgg
     2041 taccgcgggc ccgggatcca ccggatctag ataactgatc aactcctcag gtgcaggctg
     2101 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     2161 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     2221 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     2281 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     2341 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     2401 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     2461 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     2521 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     2581 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg
     2641 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2701 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2761 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     2821 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2881 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2941 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3001 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3061 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3121 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     3181 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3241 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3301 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3361 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3421 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3481 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3541 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3601 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3661 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3721 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3781 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3841 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3901 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3961 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4021 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4081 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4141 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4201 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4261 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4321 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4381 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4441 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4501 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4561 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4621 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4681 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4741 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4801 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4861 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4921 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4981 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5041 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5101 ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt cgacattgat tattgactag
     5161 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     5221 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     5281 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     5341 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     5401 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     5461 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     5521 ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcc
//
