LOCUS       sequence-360376-        5264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for transient expression of plasmodesmata-targeted mCherry
            in plants. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5264)
  AUTHORS   Han J, Ma K, Li H, Su J, Zhou L, Tang J, Zhang S, Hou Y, Chen L, Liu
            YG, Zhu Q
  TITLE     All-in-one: A Robust Fluorescent Fusion Protein Vector Toolbox for 
            Protein Localization and BiFC Analyses in Plants.
  JOURNAL   Plant Biotechnol J. 2022 Feb 18. doi: 10.1111/pbi.13790.
  PUBMED    35179286
REFERENCE   2  (bases 1 to 5264)
  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: "Plant 
            Biotechnol J. 2022 Feb 18. doi: 10.1111/pbi.13790."
COMMENT     Sequence Label: pUC19/Pd-mChe
FEATURES             Location/Qualifiers
     source          1..5264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        535..880
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     823..843
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    898..922
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     RBS             934..954
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1567..2277
                     /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(1712..1730)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1980..1999
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2303..2555
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2612..2629)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2612..2628)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2621..2643)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2837..2856)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2956..2978
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3016..3034)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3102..3206
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3207..3275,3276..4067)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSV
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3425..3444)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4238..4826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4727..4746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4980..4997
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5114..5135
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5150..5155,5156..5173,5174..5180)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5188..5204
                     /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     5193..5215
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5212..5228
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5212..5228
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggcgcgccat tactcgcatc cattctcagg ctgtctcgtc tcgtctcaga ttagcctttt
       61 caatttcaga aagaatgcta acccacagat ggttagagag gcttacgcag caggtctcat
      121 caagacgatc tacccgagca ataatctcca ggaaatcaaa taccttccca agaaggttaa
      181 agatgcagtc aaaagattca ggactaactg catcaagaac acagagaaag atatatttct
      241 caagatcaga agtactattc cagtatggac gattcaaggc ttgcttcaca aaccaaggca
      301 agtaatagag attggagtct ctaaaaaggt agttcccact gaatcaaagg ccatggagtc
      361 aaagattcaa atagaggacc taacagaact cgccgtaaag actggcgaac agttcataca
      421 gagtctctta cgactcaatg acaagaagaa aatcttcgtc aacatggtgg agcacgacac
      481 acttgtctac tccaaaaata tcaaagatac agtctcagaa gaccaaaggg caattgagac
      541 ttttcaacaa agggtaatat ccggaaacct cctcggattc cattgcccag ctatctgtca
      601 ctttattgtg aagatagtgg aaaaggaagg tggctcctac aaatgccatc attgcgataa
      661 aggaaaggcc atcgttgaag atgcctctgc cgacagtggt cccaaagatg gacccccacc
      721 cacgaggagc atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg
      781 atgtgatatc tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc
      841 ttcctctata taaggaagtt catttcattt ggagagaaca cgggggactc tagatcaaca
      901 agtttgtaca aaaaagcagg ctccgcggcc gccttgttta actttaagaa ggagcccttc
      961 accatggctg ctctggtgct ttcacttctc cttctatcct tggctggaca ttctagtgcc
     1021 tcatggtgtg tgtgtaagac agggctgagt gatacagtgc tacaggcaac cctagactat
     1081 gcttgtggaa atggagcaga ttgtaatcct actaaaccaa aacaatcttg cttcaaccct
     1141 gacaatgtta ggtctcattg caactatgca gtcaatagct tcttccaaaa gaagggtcaa
     1201 tctcctggct cttgtaattt cgatggaact gccactccca ctaactccga tcccagttat
     1261 acaggttgtg cctttcctac tagtgccagt ggctctagcg gcagcacaac tgtgacacct
     1321 ggcacaacca atccaaaagg cagcccaacg accaccacac ttcctggtag tggtaccaac
     1381 agtccttatt cagggaaccc aaccaatgga gtttttgggg gaaatagcac aggaggcacc
     1441 actgggacag ggattaaccc ggattacacg acagacagca gcgcgtttgc tctcaagaac
     1501 tcaagcaaat tgttcatctg ccttctcttg atcgcttcga gtggattctg ctctttcctg
     1561 atgctcatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     1621 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1681 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     1741 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     1801 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     1861 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     1921 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1981 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     2041 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     2101 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     2161 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2221 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaataa
     2281 ctcgagaagc tcgaatttcc ccgatcgttc aaacatttgg caataaagtt tcttaagatt
     2341 gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt acgttaagca
     2401 tgtaataatt aacatgtaat gcatgacgtt atttatgaga tgggttttat gattagagtc
     2461 ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa
     2521 ttatcgcgcg cggtgtcatc taatgttact agatcgggcc aggatacata gattaccaca
     2581 actccgagcc cttccaccgg cgcgccaatt cactggccgt cgttttacaa cgtcgtgact
     2641 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2701 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2761 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2821 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     2881 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2941 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     3001 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     3061 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3121 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3181 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3241 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3301 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3361 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3421 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3481 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3541 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3601 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3661 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3721 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3781 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3841 ataaagttgc aggaccactt ctgcgctcgg tccttccggc tggctggttt attgctgata
     3901 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3961 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4021 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4081 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4141 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4201 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4261 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4321 aagagctacc aactcttttt ccgaaggtaa ctggcttcag tagagcgcag ataccaaata
     4381 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4441 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4501 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4561 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4621 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4681 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4741 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4801 cgtcaggggg gcggagccta tggaaaaacg ctagcaacgc ggccttttta cggttcctgg
     4861 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4921 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4981 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     5041 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5101 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     5161 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     5221 gctatgacca tgattacgcc aagcttgcat gcctgcaggt cccc
//
