LOCUS       sequence-360361-        4748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for transient expression of endoplasmic reticulum-targeted
            mCherry in plants. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4748)
  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 4748)
  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/ER-mChe
FEATURES             Location/Qualifiers
     source          1..4748
                     /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             1039..1746
                     /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(1184..1202)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1452..1471
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      1787..2039
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2096..2113)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2096..2112)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2105..2127)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2321..2340)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2440..2462
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2500..2518)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2586..2690
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2691..2759,2760..3551)
                     /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(2909..2928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3722..4310
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4211..4230
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4464..4481
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4598..4619
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4634..4639,4640..4657,4658..4664)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4672..4688
                     /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     4677..4699
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4696..4712
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4696..4712
                     /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 accatgaaga gtactatctt ctttgctctc tttctagttt gtgccttcac catctcatac
     1021 ctgccttcag ccaccgctat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
     1081 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     1141 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     1201 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     1261 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     1321 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     1381 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     1441 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     1501 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     1561 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     1621 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     1681 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     1741 tacaagaaag atgaactcta ataactcgag aagctcgaat ttccccgatc gttcaaacat
     1801 ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata
     1861 atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat
     1921 gagatgggtt ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa
     1981 atatagcgcg caaactagga taaattatcg cgcgcggtgt catctaatgt tactagatcg
     2041 ggccaggata catagattac cacaactccg agcccttcca ccggcgcgcc aattcactgg
     2101 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     2161 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     2221 cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc
     2281 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     2341 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     2401 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     2461 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     2521 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     2581 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2641 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2701 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2761 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2821 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2881 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2941 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3001 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3061 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3121 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3181 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3241 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3301 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggtccttc
     3361 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3421 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3481 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3541 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3601 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3661 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3721 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3781 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3841 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3901 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3961 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4021 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4081 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4141 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4201 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4261 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgctagca
     4321 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4381 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4441 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     4501 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     4561 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     4621 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     4681 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt gcatgcctgc
     4741 aggtcccc
//
