LOCUS       sequence-360366-        4832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for transient expression of plastids-targeted mCherry in 
            plants. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4832)
  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 4832)
  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/Pt-mChe
FEATURES             Location/Qualifiers
     source          1..4832
                     /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             1135..1845
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVNHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1280..1298)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1548..1567
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      1871..2123
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2180..2197)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2180..2196)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2189..2211)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2405..2424)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2524..2546
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2584..2602)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2670..2774
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2775..2843,2844..3635)
                     /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(2993..3012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3806..4394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4295..4314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4548..4565
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4682..4703
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4718..4723,4724..4741,4742..4748)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4756..4772
                     /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     4761..4783
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4780..4796
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4780..4796
                     /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 accatggctt ctatgatatc ctcttccgct gtgacaacag tcagccgtgc ctctaggggg
     1021 caatccgccg cagtggctcc attcggcggc ctcaaatcca tgactggatt cccagtgaag
     1081 aaggtcaaca ctgacattac ttccattaca agcaatggtg gaagagtaaa gtgcatggtg
     1141 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     1201 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1261 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     1321 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa tcaccccgcc
     1381 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     1441 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     1501 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     1561 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     1621 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     1681 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     1741 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     1801 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaataact cgagaagctc
     1861 gaatttcccc gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc
     1921 cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg taataattaa
     1981 catgtaatgc atgacgttat ttatgagatg ggttttatga ttagagtccc gcaattatac
     2041 atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg
     2101 gtgtcatcta atgttactag atcgggccag gatacataga ttaccacaac tccgagccct
     2161 tccaccggcg cgccaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     2221 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     2281 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
     2341 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     2401 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     2461 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2521 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     2581 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     2641 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2701 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2761 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2821 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2881 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2941 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3001 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3061 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3121 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3181 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3241 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3301 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3361 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3421 gaccacttct gcgctcggtc cttccggctg gctggtttat tgctgataaa tctggagccg
     3481 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3541 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3601 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3661 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3721 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3781 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3841 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3901 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3961 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4021 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4081 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4141 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4201 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4261 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4321 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4381 ggagcctatg gaaaaacgct agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4441 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4501 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4561 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4621 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     4681 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     4741 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     4801 attacgccaa gcttgcatgc ctgcaggtcc cc
//
