LOCUS       sequence-391140-        4933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes mCherry-GFP1-10 in pDGB1-aR backbone for plant expression. 
            Requires pSOUP for replication in agrobacterium..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4933)
  AUTHORS   Wang JW, Squire HJ, Goh NS, Ni HM, Lien E, Wong C, Gonzalez-Grandio 
            E, Landry MP
  TITLE     Delivered complementation in planta (DCIP) enables measurement of 
            peptide-mediated protein delivery efficiency in plants.
  JOURNAL   Commun Biol. 2023 Aug 12;6(1):840. doi: 10.1038/s42003-023-05191-5.
  PUBMED    37573467
REFERENCE   2  (bases 1 to 4933)
  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: "Commun 
            Biol."; date: "2023-08-12"; volume: "6(1):840. doi"; pages: " 
            10.1038/s42003-023-05191-5"
COMMENT     Sequence Label: cytoDCIP
FEATURES             Location/Qualifiers
     source          1..4933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      672..1260
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1161..1180
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1351..1375
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        1396..1741
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1684..1704
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    1761..1816
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             1835..2542
                     /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(1980..1998)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2248..2267
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2612..2640)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     terminator      3250..3502
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3537..3555)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3744..3766
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      3893..4328
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     primer_bind     complement(4608..4627)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(4619..4933,1..501)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             join(4619..4933,1..501)
                     /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
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4692..4711)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
       61 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
      121 gtatttcgac tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
      181 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
      241 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
      301 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
      361 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
      421 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
      481 atgctcgatg agtttttcta atcactagac caatgttaca catatatact ttagattgat
      541 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
      601 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
      661 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
      721 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct tcttccgaag
      781 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
      841 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      901 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      961 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1021 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     1081 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     1141 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1201 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1261 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     1321 agatctcaaa caaacacata cagcgactta gtttacccgc caatatatcc tgtcaggatc
     1381 cgtctcatga cggagtgaga cttttcaaca aagggtaata tcgggaaacc tcctcggatt
     1441 ccattgccca gctatctgtc acttcatcga aaggacagta gaaaaggaag gtggctccta
     1501 caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcctcta ccgacagtgg
     1561 tcccaaagat ggacccccac ccacgaggaa catcgtggaa aaagaagatg ttccaaccac
     1621 gtcctcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc
     1681 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac
     1741 aggcttcttg agatccttca acaattacca acaacaacaa acaacaaaca acattacaat
     1801 tactatttac aattacagtc gactctagag gataatggtg agcaagggcg aggaggataa
     1861 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     1921 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     1981 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     2041 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     2101 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     2161 ggtgaccgtg acccaggact cctccctcca ggacggcgag ttcatctaca aggtgaagct
     2221 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     2281 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     2341 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     2401 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     2461 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     2521 cggcatggac gagctgtaca agggcggtgg cggtagcggc ggtggcagcc tgcagatgat
     2581 cgatagcaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
     2641 tgatgttaat gggcacaaat tttctgtccg tggagagggt gaaggtgatg ctacaattgg
     2701 aaaactcacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
     2761 tgtcactact ctgacctatg gtgttcaatg cttttcccgt tatccggatc acatgaaacg
     2821 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaacgca ctatatcttt
     2881 caaagatgac gggaaataca agacgcgtgc tgtcgtcaag tttgaaggtg atacccttgt
     2941 taatcgtatc gagttaaaag gtactgattt taaagaagat ggaaacattc tcggacacaa
     3001 actcgagtac aactttaact cacacaatgt atacatcacg gcagacaaac aaaagaatgg
     3061 aatcaaagct aacttcaccg ttcgccacaa cgttgaagat ggttccgttc aactagcaga
     3121 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
     3181 cctgtcgaca caaactgtcc tttcgaaaga tcccaacgaa aagtgagctt ggaatggatc
     3241 ttcgatcccg atcgttcaaa catttggcaa taaagtttct taagattgaa tcctgttgcc
     3301 ggtcttgcga cgattatcat ataatttctg ttgaattacg ttaagcatgt aataattaac
     3361 atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc gcaattatac
     3421 atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg
     3481 gtgtcatcta tgttactaga tcgggaattg ccaagctaat tcttgaagat gaaagggcct
     3541 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3601 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3661 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat gggaccgact
     3721 cgcgctctcc tgagacggat ccgttacacc acaatatatc ctgccaagat ctaattccgg
     3781 ggatcggaaa tccagaagcc cgagaggttg ccgcctttcg ggctttttct ttttcaaaaa
     3841 aaaaaattta taaaacgatc tgttgcggcc ggccgccggg ttgtgggcaa aggcgctcga
     3901 cggtgggcaa ccgcttgcgg ttgtccacgg gcggagccgg tgcgcgtagc gcattgtcca
     3961 caagccaagg gcgaccaata attgatatat atattcataa ttgaaaagct aattgaacat
     4021 actacttgct gtaactactt gccggagcga ggggtgtttg caagctgttg atctgaaagg
     4081 gctattagcg ttctcacgtg cctttttgat tagcgatttc acgtgacctt attagcgatt
     4141 tcacgtactc cgattagcga tttcacgtac cctgattagc gatttcacgt ggatagtttt
     4201 tggagcgggc cggaaagccc cgtgaatcaa ggctttgcgg ggcattagcg gtttcacgtg
     4261 gataactacc ctctatccac aggcttccgg ggataaaaaa gcccgctcga cggcgggctg
     4321 ttggatgggg atcgcctgaa tcgccccatc atccagccag aaagtgaggg agccacggtt
     4381 gatgagagct ttgttgtagg tggaccagtt ggtgattttg aacttttgct ttgccacgga
     4441 acggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa aagttcgatt
     4501 tattcaacaa agccacgttg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     4561 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     4621 gagccatatt caacgggaaa cgtcttgctc aaggccgcga ttaaattcca acatggatgc
     4681 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     4741 ccgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     4801 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     4861 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     4921 cccagggaaa aca
//
