LOCUS       sequence-367132-        5692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for transient expression of cytoplasmic foci-targeted 
            mCherry in plants. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5692)
  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 5692)
  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/CF-mChe
FEATURES             Location/Qualifiers
     source          1..5692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        535..879
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     822..842
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    897..921
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     RBS             933..953
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1995..2705
                     /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(2140..2158)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2408..2427
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2731..2983
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3040..3057)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3040..3056)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3049..3071)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3265..3284)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3384..3406
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3444..3462)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3530..3634
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3635..3703,3704..4495)
                     /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(3853..3872)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4666..5254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5155..5174
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5408..5425
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5542..5563
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5578..5583,5584..5601,5602..5608)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5616..5632
                     /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     5621..5643
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5640..5656
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5640..5656
                     /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 agaagacgtt ccaaccacgt cttcaaagca agtggattga
      781 tgtgatatct ccactgacgt aagggatgac gcacaatccc actatccttc gcaagaccct
      841 tcctctatat aaggaagttc atttcatttg gagagaacac gggggactct agatcaacaa
      901 gtttgtacaa aaaagcaggc tccgcggccg ccttgtttaa ctttaagaag gagcccttca
      961 ccagccaact ccctttacaa cctcactcaa gtccgttaga gcccatgatg atcggcgaaa
     1021 ataaaaaccg gccacatcca acgatccata tccctcaatg ggatcaaatc aacgatccaa
     1081 cggccacaat ctcttcacca ttctcttccg tcaaccttaa cagcgttaac gactacccac
     1141 actctccgtc accgtatctc gactccttcg cttctctctt ccgttacctc ccgtcaaacg
     1201 agttaacaaa cgattcagac tcatcaagtg gcgacgagtc atcaccactc accgactcat
     1261 tctcctccga cgagtttcgc atctacgagt tcaaaatccg gcgatgcgct cgaggtcgat
     1321 ctcatgattg gacggagtgt ccgttcgcac atcccggaga aaaagctcga cgacgtgatc
     1381 cgagaaagtt tcattactcc ggcaccgctt gtcctgagtt tcgtaaagga agttgtagaa
     1441 gaggtgattc gtgtgagttc tctcatggag ttttcgagtg ttggctccat ccttctcgtt
     1501 accgtactca gccgtgtaaa gacggaacta gctgccggag aagaatctgt ttcttcgctc
     1561 atacgacgga gcagttacgt gtattacctt gttcgttaga tccagatctt ggattcttct
     1621 caggattagc tacttctccg acttcgattc ttgtttctcc ttcgttttca ccaccgtcgg
     1681 aatctccgcc gctttctccg agtaccggtg aacttattgc gtcgatgagg aaaatgcaat
     1741 tgaacggagg tggttgttcg tggagttctc cgatgagatc tgcagttagg ttaccttttt
     1801 cgtcgtctct gcgtccgatt caggcggcaa cgtggccgag gataagagag tttgagatcg
     1861 aagaagctcc ggcgatggaa tttgtggaat ctgggaaaga gctgagagcg gagatgtatg
     1921 caagactcag tagagagaac tcactcggtg ggctcgttcg ctgccaccta agaatactct
     1981 acggtcacat acctatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     2041 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     2101 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     2161 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     2221 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     2281 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     2341 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     2401 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     2461 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     2521 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     2581 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     2641 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     2701 agtaataact cgagaagctc gaatttcccc gatcgttcaa acatttggca ataaagtttc
     2761 ttaagattga atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac
     2821 gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg ggttttatga
     2881 ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact
     2941 aggataaatt atcgcgcgcg gtgtcatcta atgttactag atcgggccag gatacataga
     3001 ttaccacaac tccgagccct tccaccggcg cgccaattca ctggccgtcg ttttacaacg
     3061 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     3121 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     3181 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3241 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     3301 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3361 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3421 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3481 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3541 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3601 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3661 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3721 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3781 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3841 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3901 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3961 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4021 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4081 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4141 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4201 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4261 ggaggcggat aaagttgcag gaccacttct gcgctcggtc cttccggctg gctggtttat
     4321 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4381 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4441 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4501 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4561 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4621 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4681 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4741 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4801 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4861 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4921 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4981 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5041 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5101 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5161 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5221 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgct agcaacgcgg cctttttacg
     5281 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5341 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5401 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     5461 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     5521 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     5581 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     5641 aggaaacagc tatgaccatg attacgccaa gcttgcatgc ctgcaggtcc cc
//
