LOCUS       sequence-360381-        4745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for transient expression of tonoplast-targeted mCherry in 
            plants. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4745)
  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 4745)
  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/Tp-mChe
FEATURES             Location/Qualifiers
     source          1..4745
                     /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             1048..1758
                     /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(1193..1211)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1461..1480
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      1784..2036
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2093..2110)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2093..2109)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2102..2124)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2318..2337)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2437..2459
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2497..2515)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2583..2687
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2688..2756,2757..3548)
                     /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(2906..2925)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3719..4307
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4208..4227
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4461..4478
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4595..4616
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4631..4636,4637..4654,4655..4661)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4669..4685
                     /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     4674..4696
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4693..4709
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4693..4709
                     /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 accatgtcgc agtgcgttga cggtatcaag catttatgca cctctgtttt agggtgtttt
     1021 gatcttgatt tgtataaaca atctggtatg gtgagcaagg gcgaggagga taacatggcc
     1081 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     1141 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     1201 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     1261 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     1321 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     1381 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     1441 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     1501 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     1561 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     1621 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     1681 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     1741 gacgagctgt acaagtaata actcgagaag ctcgaatttc cccgatcgtt caaacatttg
     1801 gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt
     1861 tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag
     1921 atgggtttta tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata
     1981 tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctaatgttac tagatcgggc
     2041 caggatacat agattaccac aactccgagc ccttccaccg gcgcgccaat tcactggccg
     2101 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     2161 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     2221 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     2281 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     2341 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     2401 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     2461 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     2521 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     2581 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2641 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2701 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2761 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2821 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2881 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     2941 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3001 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3061 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3121 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3181 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3241 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3301 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gtccttccgg
     3361 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3421 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3481 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3541 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3601 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3661 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3721 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3781 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3841 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3901 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3961 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4021 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4081 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4141 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4201 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4261 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gctagcaacg
     4321 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4381 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4441 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     4501 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     4561 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     4621 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     4681 aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg
     4741 tcccc
//
