LOCUS       sequence-197538-        6276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent tagging. Plant expression of YFPc-RabG3b-DN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6276)
  TITLE     Robatzek Lab plasmids
REFERENCE   2  (bases 1 to 6276)
  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"
COMMENT     Sequence Label: 35S:YFPc-RabG3b-DN
FEATURES             Location/Qualifiers
     source          1..6276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    917..941
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(1035..1287)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1307..1858)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(1900..2079)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2548..2892
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2836..2856
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2894..2910
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2895..2911
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2926..3177
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     3114..3135
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    3190..3214
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3863..3887)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3897..3923
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3936..3959
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    4252..4276
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4388..5097
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5170..5261
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5262..5330,5331..6122)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5480..5499)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      661 acatggactc tagctagagg atcacaggca gcaacgctct gtcatcgtta caatcaacat
      721 gctaccctcc gcgagatcat ccgtgtttca aacccggcag cttagttgcc gttcttccga
      781 atagcatcgg taacatgagc aaagtctgcc gccttacaac ggctctcccg ctgacgccgt
      841 cccggactga tgggctgcct gtatcgagtg gtgattttgt gccgagctgc cggtcgggga
      901 gctgttggct ggctggtggc aggatatatt gtggtgtaaa caaattgacg cttagacaac
      961 ttaataacac attgcggacg tttttaatga tcgaatacta acgtctctac cagatatcag
     1021 cttgcatgcc ggtcgatcta gtaacataga tgacaccgcg cgcgataatt tatcctagtt
     1081 tgcgcgctat attttgtttt ctatcgcgta ttaaatgtat aattgcggga ctctaatcat
     1141 aaaaacccat ctcataaata acgtcatgca ttacatgtta attattacat gcttaacgta
     1201 attcaacaga aattatatga taatcatcgc aagaccggca acaggattca atcttaagaa
     1261 actttattgc caaatgtttg aacgatctgc ttgactctag gggtcatcag atttcggtga
     1321 cgggcaggac cggacggggc ggcaccggca ggctgaagtc cagctgccag aaacccacgt
     1381 catgccagtt cccgtgcttg aagccggccg cccgcagcat gccgcggggg gcatatccga
     1441 gcgcctcgtg catgcgcacg ctcgggtcgt tgggcagccc gatgacagcg accacgctct
     1501 tgaagccctg tgcctccagg gacttcagca ggtgggtgta gagcgtggag cccagtcccg
     1561 tccgctggtg gcggggggag acgtacacgg tcgactcggc cgtccagtcg taggcgttgc
     1621 gtgccttcca gggacccgcg taggcgatgc cggcgacctc gccgtccacc tcggcgacga
     1681 gccagggata gcgctcccgc agacggacga ggtcgtccgt ccactcctgc ggttcctgcg
     1741 gctcggtacg gaagttgacc gtgcttgtct cgatgtagtg gttgacgatg gtgcagaccg
     1801 ccggcatgtc cgcctcggtg gcacggcgga tgtcggccgg gcgtcgttct gggctcatgg
     1861 tagatcccct cgatcgagtt gagagtgaat atgagactct aattggatac cgaggggaat
     1921 ttatggaacg tcagtggagc atttttgaca agaaatattt gctagctgat agtgacctta
     1981 ggcgactttt gaacgcgcaa taatggtttc tgacgtatgt gcttagctca ttaaactcca
     2041 gaaacccgcg gctcagtggc tccttcaacg ttgcggttct gtcagttcca aacgtaaaac
     2101 ggcttgtccc gcgtcatcgg cgggggtcat aacgtgactc ccttaattct ccgctcatga
     2161 tcagattgtc gtttcccgcc ttcggtttgg gcgcgcccgg ttcagaagac cagagggcta
     2221 ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct cggattccat tgcccagcta
     2281 tctgtcactt catcgaaagg acagtagaaa aggaagatgg cttctacaaa tgccatcatt
     2341 gcgataaagg aaaggctatc gttcaagatg cctctaccga cagtggtccc aaagatggac
     2401 ccccacccac gaggaacatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag
     2461 tggattgatg tgatacatgg tggagcacga cactctcgtc tactccaaga atatcaaaga
     2521 tacagtctca gaagaccaga gggctattga gacttttcaa caaagggtaa tatcgggaaa
     2581 cctcctcgga ttccattgcc cagctatctg tcacttcatc gaaaggacag tagaaaagga
     2641 agatggcttc tacaaatgcc atcattgcga taaaggaaag gctatcgttc aagatgcctc
     2701 taccgacagt ggtcccaaag atggaccccc acccacgagg aacatcgtgg aaaaagaaga
     2761 cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat atctccactg acgtaaggga
     2821 tgacgcacaa tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca
     2881 tttggagagg acctcgaggt cgacggtatc gataagctta taatggacaa gcagaagaac
     2941 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     3001 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3061 tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3121 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggga
     3181 gtccccatca caagtttgta caaaaaagca ggctccgcgg ccgccccctt caccatgtcg
     3241 acgcgaagac gaactttact caaagttata attcttggag acagcggggt tggcaaaaac
     3301 tcgttgatga atcaatatgt gaataacaag tttagtcaac agtacaaagc tacgatcgga
     3361 gctgattttg tcactaagga gcttcaaatt gatgacaggc ttgtcacatt gcaaatatgg
     3421 gacactgctg ggcaagagag gtttcaaagt cttggtgttg ctttctatag aggtgcagat
     3481 tgttgtgttc ttgtctatga tgtgaatcac ttgaagtcat ttgaatctct cgacaattgg
     3541 cacaacgagt ttcttacacg ggctagtcca cgtgacccaa tggcattccc ttttatactt
     3601 cttggtaata aggttgatat tgatggagga aatagccgag tggtatctga gaagaaggct
     3661 agagaatggt gtgctgaaaa gggaaacata gtctatttcg agacatcggc taaagaagat
     3721 tacaatgtcg atgactcctt cttgtgcatc acaaaacttg cccttgcaaa tgaacgcgac
     3781 caagatatat atttccagcc agatactggt tcggtgcctg agcaaagagg aggttgtgct
     3841 tgctgaaagg gtgggcgcgc cgacccagct ttcttgtaca aagtggtgat gggggttatc
     3901 catacgatgt tccagattat gctgtcggcg ccggttggtc tcatcctcaa tttgaaaaat
     3961 aagtctagag tccgcaaaaa tcaccagtct ctctctacaa atctatctct ctctattttt
     4021 ctccagaata atgtgtgagt agttcccaga taagggaatt agggttctta tagggtttcg
     4081 ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta
     4141 tcaataaaat ttctaattcc taaaaccaaa atccagtgac cgggcggccg ccaccgcggt
     4201 ggagggggat cagattgtcg tttcccgcct tcagtttaaa ctatcagtgt ttgacaggat
     4261 atattggcgg gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag
     4321 ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgta catcaccgac gagcaaggca
     4381 agaccgagcg cctttccgac gctcaccggg ctggttgccc tcgccgctgg gctggcggcc
     4441 gtctatggcc ctgcaaacgc gccagaaacg ccgtcgaagc cgtgtgcgag acaccggccg
     4501 ccggcgttgt ggatacctcg cggaaaactt ggccctcact gacagatgag gggcggacgt
     4561 tgacacttga ggggccgact cacccggcgc ggcgttgaca gatgaggggc aggctcgatt
     4621 tcggccggcg acgtggagct ggccagcctc gcaaatcggc gaaaacgcct gattttacgc
     4681 gagtttccca cagatgatgt ggacaagcct ggggataagt gccctgcggt attgacactt
     4741 gaggggcgcg actactgaca gatgaggggc gcgatccttg acacttgagg ggcagagtgc
     4801 tgacagatga ggggcgcacc tattgacatt tgaggggctg tccacaggca gaaaatccag
     4861 catttgcaag ggtttccgcc cgtttttcgg ccaccgctaa cctgtctttt aacctgcttt
     4921 taaaccaata tttataaacc ttgtttttaa ccagggctgc gccctgtgcg cgtgaccgcg
     4981 cacgccgaag gggggtgccc ccccttctcg aaccctcccg gcccgctaac gcgggcctcc
     5041 catcccccca ggggctgcgc ccctcggccg cgaacggcct caccccaaaa atggcagcgc
     5101 tggcagtcct tgccattgcc gggatcgggg cagtaacggg atgggcgatc agcccgaaag
     5161 ctacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5221 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5281 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5341 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5401 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5461 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5521 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5581 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5641 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5701 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5761 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5821 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5881 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5941 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6001 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6061 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6121 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6181 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6241 agttttcgtt ccactgagcg tcagaccccg tagaaa
//
