LOCUS       sequence-367124-        5303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for transient expression of plasma membrane-targeted 
            mCherry in plants. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5303)
  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 5303)
  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-PM
FEATURES             Location/Qualifiers
     source          1..5303
                     /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             964..1671
                     /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(1109..1127)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1377..1396
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2342..2594
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2651..2668)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2651..2667)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2660..2682)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2876..2895)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2995..3017
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3055..3073)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3141..3245
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3246..3314,3315..4106)
                     /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(3464..3483)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4277..4865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4766..4785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5019..5036
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5153..5174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5189..5194,5195..5212,5213..5219)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5227..5243
                     /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     5232..5254
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5251..5267
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5251..5267
                     /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 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     1021 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     1081 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     1141 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     1201 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     1261 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     1321 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     1381 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     1441 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     1501 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     1561 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     1621 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gatggcgtcc
     1681 agcgcctccc ggttcatcaa gtgcgtcacg gtgggcgacg tagccgtggg caagacctgc
     1741 atgctcatct gctacaccag caacaagttc cccactgact acataccaac ggtgttcgac
     1801 aatttcagcg caaacgttgt tgtggacagc accacggtga atctgggcct ctgggatact
     1861 gctgggcaag aggattacaa ccggctcagg cctctgagct atcgtggtgc tgatgttttc
     1921 gtgcttgcct tctctcttgt gagccgagct agctatgaaa atattatgaa gaagtggata
     1981 ccggagctac agcattatgc acccggcgtg cccattgtgt tggttggcac aaaattggat
     2041 ctccgtgaag acaagcacta cttgttggac catcctggca tgatacctgt taccacagca
     2101 cagggtgaag aacttcgaaa acaaataggt gctgcttatt acattgagtg cagctcaaag
     2161 acacaacaga atgtcaaagg tgtgtttgat gctgctatca aggtagtaat ccagcctcca
     2221 actaagcaga gggaaaagaa gaaaaagaaa tcacgacaag gatgctctat gatgaacatg
     2281 ttccgtggaa ggaaaatgtc atgcttcaaa tcctaataac tcgagaagct cgaatttccc
     2341 cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc
     2401 gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg
     2461 catgacgtta tttatgagat gggttttatg attagagtcc cgcaattata catttaatac
     2521 gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct
     2581 aatgttacta gatcgggcca ggatacatag attaccacaa ctccgagccc ttccaccggc
     2641 gcgccaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     2701 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     2761 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     2821 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     2881 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     2941 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3001 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     3061 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3121 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3181 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3241 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3301 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3361 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3421 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3481 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3541 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3601 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3661 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3721 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3781 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3841 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3901 tgcgctcggt ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3961 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4021 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4081 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4141 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4201 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4261 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4321 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4381 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4441 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4501 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4561 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4621 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4681 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4741 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4801 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4861 ggaaaaacgc tagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4921 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4981 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5041 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5101 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     5161 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     5221 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     5281 agcttgcatg cctgcaggtc ccc
//
