LOCUS       sequence-360373-        5750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for transient expression of cytoskeleton-targeted mCherry 
            in plants. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5750)
  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 5750)
  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-Cs
FEATURES             Location/Qualifiers
     source          1..5750
                     /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      2789..3041
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3098..3115)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3098..3114)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3107..3129)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3323..3342)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3442..3464
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3502..3520)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3588..3692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3693..3761,3762..4553)
                     /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(3911..3930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4724..5312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5213..5232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5600..5621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5636..5641,5642..5659,5660..5666)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5674..5690
                     /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     5679..5701
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5698..5714
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5698..5714
                     /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 ggatcctctt
     1681 gaaagagctg aattggttct cagtcatgca gagagaatga actgcaagcg atacttgact
     1741 gcagaggaga tcgttgaagg atcttcaact ttgaatcttg cctttgtggc acaaatcttc
     1801 catgaaagga atggtctaaa caaggatggt aagtatgcat ttgcggagat gatgaccgaa
     1861 gacgtagaga cttgtagaga tgaacgatgc taccggctat ggatcaacag cctcgggatt
     1921 gacagttatg tcaataatgt atttgaagat gttagaaatg gatggattct tcttgaggtt
     1981 cttgacaagg tctctcccag ttcagtcaac tggaagcatg cttccaaacc accgattaag
     2041 atgccgttta gaaaagtaga gaactgcaat caagtcataa agatcgggaa acagctaaaa
     2101 ttctcacttg taaatgtagc tggaaatgac atagttcaag ggaataagaa gctcattctt
     2161 ggtctcttat ggcagttgat gagattccat atgctccaac ttctcaagag tctcagatct
     2221 cggacactag gtaaagagat gactgatgca gatatcctca gctgggccaa caggaaagta
     2281 agaacaatgg gacgaaaatt gcaaatcgag agtttcaagg acaagagtct atcgagtggg
     2341 ttattcttcc tcaaccttct atgggcggtt gaaccaagag ttgtgaactg gaatcttgtc
     2401 accaagggtg aaacagatga tgagaagagg ttgaatgcta catacatagt tagtgtcgca
     2461 agaaagctcg gttgttcggt tttcttgtta cctgaagata tcgtggaggt gaatcagaag
     2521 atgatcctaa ttttaacggc aagtataatg tactggagtc ttcagagaca ttcacgggag
     2581 agttcagatt cgtcgtcaac tcagagcacc acaacgacgt gcaccagcac agcctcgtcc
     2641 cctgccccat ctgtcacaga agaggaggaa gtctcctcat tgagcggtga agtcacgagc
     2701 ttggccgttg gtgatgcggt ttctgaaatc accacggtct cagaggaagc atcaatcgaa
     2761 taataactcg agaagctcga atttccccga tcgttcaaac atttggcaat aaagtttctt
     2821 aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt
     2881 taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg ttttatgatt
     2941 agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg cgcaaactag
     3001 gataaattat cgcgcgcggt gtcatctaat gttactagat cgggccagga tacatagatt
     3061 accacaactc cgagcccttc caccggcgcg ccaattcact ggccgtcgtt ttacaacgtc
     3121 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     3181 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     3241 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3301 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     3361 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3421 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3481 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3541 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3601 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3661 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3721 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3781 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3841 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3901 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3961 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4021 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4081 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4141 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4201 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4261 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4321 aggcggataa agttgcagga ccacttctgc gctcggtcct tccggctggc tggtttattg
     4381 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4441 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4501 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4561 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4621 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4681 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4741 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4801 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4861 caaatactgt tcttctagtg tagccgtagt taggccacca cctcaagaac tctgtagcac
     4921 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4981 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5041 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5101 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5161 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5221 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5281 gatgctcgtc aggggggcgg agcctatgga aaaacgctag caacgcggcc tttttacggt
     5341 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5401 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5461 agcgcagtga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     5521 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     5581 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     5641 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     5701 gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcccc
//
