LOCUS       sequence-361246-        4444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of OEP7-mCherry in plant cell (Outer envelope 
            membrane of plastid/chloroplast).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4444)
  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 4444)
  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-OEP7-mCherry
FEATURES             Location/Qualifiers
     source          1..4444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..35)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(189..777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(269..288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(935..1750)
                     /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     1658..1677
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1742..1761
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1935..1953
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1991..2013)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2113..2132
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2326..2348
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2340..2357
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2341..2357
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2367..2386
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2367..2385
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2462..2806
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2749..2769
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    2826..2881
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     protein_bind    2925..2949
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3102..3809
                     /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(3247..3265)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3515..3534
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(3811..3835)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      3880..4130
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4195..4213)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4196..4213)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4231..4247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4231..4247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4244..4266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4255..4271
                     /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(4279..4285,4286..4303,4304..4309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4324..4345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
       61 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
      121 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
      181 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
      241 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
      301 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
      361 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
      421 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
      481 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      541 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      601 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      661 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      721 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      781 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      841 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      901 agttttaaat caatctaaag tatatatgtt ttaattagaa aaactcatcg agcatcaaat
      961 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
     1021 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
     1081 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
     1141 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaatagct
     1201 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
     1261 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat
     1321 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
     1381 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
     1441 tccctgggat cgcagaggtg agtaaccatg catcatcagg agtacggata aaatgcttga
     1501 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
     1561 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
     1621 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
     1681 ataaatcagc atccatgttg gaatttaatc gcggccttga gcaagacgtt tcccgttgaa
     1741 tatggctcat aacacccctt gtattactgt ttatgtaagc agacataagc atttatcagg
     1801 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     1861 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     1921 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     1981 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     2041 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     2101 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     2161 taccgcacag atgcgtaagg agaaaatatt actacaggcg ccattcgcca ttcaggctgc
     2221 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     2281 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     2341 gtaaaacgac ggccagtgag ctagtgtaat acgactcact atagggcgcg gccgcagaat
     2401 tcgagctcgg tacccgggat ctcgaggcca gatctcccgg gctcagaaga ccgagggcta
     2461 ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct cggattccat tgcccagcta
     2521 tctgtcactt atcgaaagga cagtagaaaa ggaaggtggc tcctacaaat gccatcattg
     2581 cgataaagga aaggctatcg ttcaagatgc ctctaccgac agtggtccca aagatggacc
     2641 cccacccacg aggaacatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt
     2701 ggattgatgt gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca
     2761 agacccttcc tctatataag gaagttcatt tcatttggag aggacaggct tcttgagatc
     2821 cttcaacaat taccaacaac aacaaacaac aaacaacatt acaattacta tttacaatta
     2881 cagaattcga gctcggtacc aagcttggag gctcaggagg ctccacaagt ttgtacaaaa
     2941 aagcaggctc catgggaaaa acttcgggag cgaaacaggc gactgtggtg gtcgcagcga
     3001 tggcgttagg atggttagcc atagagatcg ctttcaagcc tttcctcgat aaattccgct
     3061 cctcaatcga caaatctgac ccaaccaaag accccgatga catggtgagc aagggcgagg
     3121 aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
     3181 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     3241 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
     3301 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
     3361 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     3421 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
     3481 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
     3541 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     3601 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
     3661 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
     3721 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
     3781 ccaccggcgg catggacgag ctgtacaagg acccagcttt cttgtacaaa gtggttggag
     3841 gctcaggagg ctccccatgg gctagcacta gtatctagaa tcgttcaaac atttggcaat
     3901 aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt
     3961 tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg
     4021 tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc
     4081 gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat gcggccgcaa
     4141 ttgtggatcc gctctagagt cgacctgcag gcatgcaagc ttgcggccgc gtattctata
     4201 gtgtcaccta aatagcatgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4261 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4321 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4381 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4441 tatt
//
