LOCUS       sequence-382265-        4324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of mCherry-NLS in plant cell (Nucleus).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4324)
  AUTHORS   Ichikawa S, Kato S, Fujii Y, Ishikawa K, Numata K, Kodama Y
  TITLE     Organellar Glue: A Molecular Tool to Artificially Control 
            Chloroplast-Chloroplast Interactions.
  JOURNAL   ACS Synth Biol. 2022 Oct 21;11(10):3190-3197. doi: 
            10.1021/acssynbio.2c00367. Epub 2022 Sep 30.
  PUBMED    36178266
REFERENCE   2  (bases 1 to 4324)
  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"; doi: 
            "10.1021/acssynbio.2c00367"; journalName: "ACS Synth Biol"; date: 
            "2022-10-21- 21"; volume: "11"; issue: "10"; pages: "3190-3197"
COMMENT     Sequence Label: pGWT35S-mCherry-NLS
FEATURES             Location/Qualifiers
     source          1..4324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     405..424
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        405..423
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        500..844
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     787..807
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    864..919
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     protein_bind    963..987
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             990..1697
                     /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(1135..1153)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1403..1422
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1704..1724
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(1729..1753)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1798..2048
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(2113..2131)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2114..2131)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2149..2165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2149..2165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2162..2184)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2173..2189
                     /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(2197..2203,2204..2221,2222..2227))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2242..2263
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2380..2397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2551..3139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2631..2650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3297..4112)
                     /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     4020..4039
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4104..4123
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     4297..4315
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaatattac tacaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgagct agtgtaatac gactcactat
      421 agggcgcggc cgcagaattc gagctcggta cccgggatct cgaggccaga tctcccgggc
      481 tcagaagacc gagggctatt gagacttttc aacaaagggt aatatcggga aacctcctcg
      541 gattccattg cccagctatc tgtcacttat cgaaaggaca gtagaaaagg aaggtggctc
      601 ctacaaatgc catcattgcg ataaaggaaa ggctatcgtt caagatgcct ctaccgacag
      661 tggtcccaaa gatggacccc cacccacgag gaacatcgtg gaaaaagaag acgttccaac
      721 cacgtcttca aagcaagtgg attgatgtga tatctccact gacgtaaggg atgacgcaca
      781 atcccactat ccttcgcaag acccttcctc tatataagga agttcatttc atttggagag
      841 gacaggcttc ttgagatcct tcaacaatta ccaacaacaa caaacaacaa acaacattac
      901 aattactatt tacaattaca gaattcgagc tcggtaccaa gcttggaggc tcaggaggct
      961 ccacaagttt gtacaaaaaa gcaggcttca tggtgagcaa gggcgaggag gataacatgg
     1021 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
     1081 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     1141 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca
     1201 tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
     1261 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
     1321 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     1381 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     1441 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga
     1501 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
     1561 ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg
     1621 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
     1681 tggacgagct gtacaagtcg ggacctaaga agaagcgcaa ggtgtgagac ccagctttct
     1741 tgtacaaagt ggttggaggc tcaggaggct ccccatgggc tagcactagt atctagaatc
     1801 gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga
     1861 ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga
     1921 cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga
     1981 tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt
     2041 tactagatgc ggccgcaatt gtggatccgc tctagagtcg acctgcaggc atgcaagctt
     2101 gcggccgcgt attctatagt gtcacctaaa tagcatggcg taatcatggt catagctgtt
     2161 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2221 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2281 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2341 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2401 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2461 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2521 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2581 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2641 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2701 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2761 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2821 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2881 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2941 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3001 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3061 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3121 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3181 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3241 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg ttttaattag
     3301 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     3361 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     3421 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     3481 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     3541 tccggtgaga atggcaatag cttatgcatt tctttccaga cttgttcaac aggccagcca
     3601 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     3661 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     3721 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     3781 tctaatacct ggaatgctgt tttccctggg atcgcagagg tgagtaacca tgcatcatca
     3841 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     3901 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     3961 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     4021 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctt
     4081 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     4141 gcagacataa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4201 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4261 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     4321 cgtc
//
