LOCUS       sequence-323961-        5608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry2 fused with an ER-targeting sequence of 
            calreticulin and the ER retrieval sequence, KDEL, under the control 
            of CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5608)
  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 5608)
  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-ER
FEATURES             Location/Qualifiers
     source          1..5608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        12..391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        393..668
                     /label=chicken beta-actin promoter
     intron          669..1685
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1609..1631
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1693..1712
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1821..2528
                     /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(1966..1984)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2637..2656)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2702..2757
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2756..2775)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3118..3134)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3118..3134)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3131..3153)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3142..3158
                     /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(3166..3172,3173..3190,3191..3196))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3211..3232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3291..3486
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3337..3472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3492..3626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3529..3548)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3583..3602
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3694..3711)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3865..4453)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3945..3964)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4624..5415,5416..5484))
                     /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     5247..5266
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5485..5589)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgggt cgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
       61 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      121 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      181 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      241 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      301 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      361 catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac
      421 tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt
      481 ttgtgcagcg atgggggcgn nggggggggg ggcgcgcgcc aggcggggcg gggcggggcg
      541 aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc
      601 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
      661 ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc
      721 gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc
      781 cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc
      841 tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg
      901 gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct
      961 gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg
     1021 cggccggggg cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc
     1081 ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc
     1141 ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta
     1201 cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg
     1261 gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga
     1321 gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg
     1381 agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc
     1441 gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg
     1501 gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg
     1561 gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct
     1621 ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc
     1681 tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagctagcc
     1741 taccggtcgc caccatgctg ctatccgtgc cgttgctgct cggcctcctc ggcctggccg
     1801 tcgccgaccg gtcgcacacc atggtgagca agggcgagga ggataacatg gccatcatca
     1861 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
     1921 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     1981 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     2041 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     2101 agggcttcaa ttgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     2161 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     2221 tcccctccga cggccccgta atgcagtgtc gtaccatggg ctgggaggcc tccactgagc
     2281 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     2341 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     2401 tgcccggcgc ctacaacgtc gacatcaagt tggacatcct ttcccacaac gaggactaca
     2461 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     2521 tgtacaagag atcgtacaag aaggacgagc tgtaataaaa ctcctcaggt gcaggctgcc
     2581 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     2641 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     2701 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     2761 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     2821 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     2881 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     2941 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     3001 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     3061 gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc
     3121 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3181 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3241 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat
     3301 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3361 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3421 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3481 tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3541 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3601 tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag
     3661 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3721 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3781 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3841 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3901 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3961 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4021 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4081 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4141 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4201 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4261 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4321 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4381 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4441 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4501 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4561 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4621 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4681 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4741 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4801 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4861 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4921 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4981 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5041 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5101 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5161 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5221 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5281 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5341 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5401 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5461 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5521 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5581 ggttccgcgc acatttcccc gaaaagtg
//
