LOCUS       sequence-367134-        4661 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for transient expression of cell nucleus-, 
            cytoplasm-targeted mCherry in plants. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4661)
  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 4661)
  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/mChe
FEATURES             Location/Qualifiers
     source          1..4661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     381..398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    515..536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(551..556,557..574,575..581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    589..605
                     /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     594..616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     613..629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     613..629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1200..1545
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1488..1508
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    1563..1587
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     RBS             1599..1619
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1629..2339
                     /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(1774..1792)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2042..2061
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2365..2617
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2674..2691)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2674..2690)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2683..2705)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2899..2918)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3018..3040
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3078..3096)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3164..3268
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3269..3337,3338..4129)
                     /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(3487..3506)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4300..4661,1..227)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
       61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gctagcaacg
      241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      541 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      601 aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg
      661 tccccggcgc gccattactc gcatccattc tcaggctgtc tcgtctcgtc tcagattagc
      721 cttttcaatt tcagaaagaa tgctaaccca cagatggtta gagaggctta cgcagcaggt
      781 ctcatcaaga cgatctaccc gagcaataat ctccaggaaa tcaaatacct tcccaagaag
      841 gttaaagatg cagtcaaaag attcaggact aactgcatca agaacacaga gaaagatata
      901 tttctcaaga tcagaagtac tattccagta tggacgattc aaggcttgct tcacaaacca
      961 aggcaagtaa tagagattgg agtctctaaa aaggtagttc ccactgaatc aaaggccatg
     1021 gagtcaaaga ttcaaataga ggacctaaca gaactcgccg taaagactgg cgaacagttc
     1081 atacagagtc tcttacgact caatgacaag aagaaaatct tcgtcaacat ggtggagcac
     1141 gacacacttg tctactccaa aaatatcaaa gatacagtct cagaagacca aagggcaatt
     1201 gagacttttc aacaaagggt aatatccgga aacctcctcg gattccattg cccagctatc
     1261 tgtcacttta ttgtgaagat agtggaaaag gaaggtggct cctacaaatg ccatcattgc
     1321 gataaaggaa aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa agatggaccc
     1381 ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg
     1441 gattgatgtg atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa
     1501 gacccttcct ctatataagg aagttcattt catttggaga gaacacgggg gactctagat
     1561 caacaagttt gtacaaaaaa gcaggctccg cggccgcctt gtttaacttt aagaaggagc
     1621 ccttcaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     1681 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     1741 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     1801 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     1861 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     1921 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     1981 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     2041 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     2101 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     2161 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     2221 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     2281 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaat
     2341 aactcgagaa gctcgaattt ccccgatcgt tcaaacattt ggcaataaag tttcttaaga
     2401 ttgaatcctg ttgccggtct tgcgatgatt atcatataat ttctgttgaa ttacgttaag
     2461 catgtaataa ttaacatgta atgcatgacg ttatttatga gatgggtttt atgattagag
     2521 tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata
     2581 aattatcgcg cgcggtgtca tctaatgtta ctagatcggg ccaggataca tagattacca
     2641 caactccgag cccttccacc ggcgcgccaa ttcactggcc gtcgttttac aacgtcgtga
     2701 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2761 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2821 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2881 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     2941 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3001 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3061 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3121 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3181 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3241 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3301 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3361 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3421 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3481 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3541 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3601 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3661 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3721 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3781 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3841 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3901 ggataaagtt gcaggaccac ttctgcgctc ggtccttccg gctggctggt ttattgctga
     3961 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4021 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4081 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4141 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4201 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4261 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4321 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4381 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4441 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4501 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4561 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4621 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc t
//
