LOCUS       sequence-361122-        4297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of mCherry in plant cell (Cytosol).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4297)
  AUTHORS   Osaki Y, Kodama Y
  TITLE     Particle bombardment and subcellular protein localization analysis 
            in the aquatic plant Egeria densa.
  JOURNAL   PeerJ. 2017 Sep 7;5:e3779. doi: 10.7717/peerj.3779. eCollection 
            2017.
  PUBMED    28894649
REFERENCE   2  (bases 1 to 4297)
  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: "PeerJ."; 
            date: "2017-09-7"; volume: "5:e3779. doi"; pages: " 
            10.7717/peerj.3779. eCollection 2017"
COMMENT     Sequence Label: pGWT35S-mCherry
FEATURES             Location/Qualifiers
     source          1..4297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     371..388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    505..526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(541..546,547..564,565..571)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    579..595
                     /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)."
     primer_bind     584..606
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     603..619
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     603..619
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        637..655
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     637..654
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(720..970)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    1015..1039
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1041..1751)
                     /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(1319..1338)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1588..1606
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    complement(1754..1778)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1822..1877
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     promoter        complement(1897..2241)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1934..1954)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(2317..2336)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2318..2336)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2346..2363)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2346..2362)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2355..2377)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2571..2590)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2690..2712
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2750..2768)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2942..2961)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2953..3768
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTSAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3026..3045)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3926..4297,1..217)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
       61 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
      241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
      301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
      361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
      421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
      481 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      541 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      601 cacaggaaac agctatgacc atgattacgc catgctattt aggtgacact atagaatacg
      661 cggccgcaag cttgcatgcc tgcaggtcga ctctagagcg gatccacaat tgcggccgca
      721 tctagtaaca tagatgacac cgcgcgcgat aatttatcct agtttgcgcg ctatattttg
      781 ttttctatcg cgtattaaat gtataattgc gggactctaa tcataaaaac ccatctcata
      841 aataacgtca tgcattacat gttaattatt acatgcttaa cgtaattcaa cagaaattat
      901 atgataatca tcgcaagacc ggcaacagga ttcaatctta agaaacttta ttgccaaatg
      961 tttgaacgat tctagatact agtgctagcc catggggagc ctcctgagcc tccaaccact
     1021 ttgtacaaga aagctgggtc tcacttgtac agctcgtcca tgccgccggt ggagtggcgg
     1081 ccctcggcgc gttcgtactg ttccacgatg gtgtagtcct cgttgtggga ggtgatgtcc
     1141 aacttgatgt tgacgttgta ggcgccgggc agctgcacgg gcttcttggc cttgtaggtg
     1201 gtcttgacct cagcgtcgta gtggccgccg tccttcagct tcagcctctg cttgatctcg
     1261 cccttcaggg cgccgtcctc ggggtacatc cgctcggagg aggcctccca gcccatggtc
     1321 ttcttctgca ttacggggcc gtcggagggg aagttggtgc cgcgcagctt caccttgtag
     1381 atgaactcgc cgtcctgcag ggaggagtcc tgggtcacgg tcaccacgcc gccgtcctcg
     1441 aagttcatca cgcgctccca cttgaagccc tcggggaagg acagcttcaa gtagtcgggg
     1501 atgtcggcgg ggtgcttcac gtaggccttg gagccgtaca tgaactgagg ggacaggatg
     1561 tcccaggcga agggcagggg gccacccttg gtcaccttca gcttggcggt ctgggtgccc
     1621 tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact cgtggccgtt cacggagccc
     1681 tccatgtgca ccttgaagcg catgaactcc ttgatgatgg ccatgttatc ctcctcgccc
     1741 ttgctcacca tggagcctgc ttttttgtac aaacttgtgg agcctcctga gcctccaagc
     1801 ttggtaccga gctcgaattc tgtaattgta aatagtaatt gtaatgttgt ttgttgtttg
     1861 ttgttgttgg taattgttga aggatctcaa gaagcctgtc ctctccaaat gaaatgaact
     1921 tccttatata gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc
     1981 agtggagata tcacatcaat ccacttgctt tgaagacgtg gttggaacgt cttctttttc
     2041 cacgatgttc ctcgtgggtg ggggtccatc tttgggacca ctgtcggtag aggcatcttg
     2101 aacgatagcc tttcctttat cgcaatgatg gcatttgtag gagccacctt ccttttctac
     2161 tgtcctttcg ataagtgaca gatagctggg caatggaatc cgaggaggtt tcccgatatt
     2221 accctttgtt gaaaagtctc aatagccctc ggtcttctga gcccgggaga tctggcctcg
     2281 agatcccggg taccgagctc gaattctgcg gccgcgccct atagtgagtc gtattacact
     2341 agctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     2401 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     2461 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgta gtaatatttt
     2521 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     2581 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     2641 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     2701 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     2761 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     2821 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2881 tatccgctca tgagacaata accctgataa atgcttatgt ctgcttacat aaacagtaat
     2941 acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctcaaggcc gcgattaaat
     3001 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     3061 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     3121 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     3181 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     3241 ctcacctctg cgatcccagg gaaaacagca ttccaggtat tagaagaata tcctgattca
     3301 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     3361 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     3421 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     3481 caagtctgga aagaaatgca taagctattg ccattctcac cggattcagt cgtcactcat
     3541 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     3601 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     3661 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     3721 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatt aaaacatata
     3781 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3841 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3901 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3961 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4021 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4081 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4141 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4201 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4261 cacagcccag cttggagcga acgacctaca ccgaact
//
