LOCUS       sequence-197536-        6243 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent tagging. Plant expression of YFPc-RabG3b-CA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6243)
  TITLE     Robatzek Lab plasmids
REFERENCE   2  (bases 1 to 6243)
  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-CA
FEATURES             Location/Qualifiers
     source          1..6243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      23..611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     512..531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    923..947
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(1041..1293)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1313..1864)
                     /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(1906..2085)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2554..2898
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2842..2862
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2900..2916
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2901..2917
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2932..3183
                     /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     3120..3141
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    3196..3220
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3836..3860)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3870..3896
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3909..3932
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    4225..4249
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4361..5070
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5143..5234
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5235..5303,5304..6095)
                     /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(5453..5472)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
       61 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      661 ttgctcacat ggactctagc tagaggatca caggcagcaa cgctctgtca tcgttacaat
      721 caacatgcta ccctccgcga gatcatccgt gtttcaaacc cggcagctta gttgccgttc
      781 ttccgaatag catcggtaac atgagcaaag tctgccgcct tacaacggct ctcccgctga
      841 cgccgtcccg gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt
      901 cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta
      961 gacaacttaa taacacattg cggacgtttt taatgatcga atactaacgt ctctaccaga
     1021 tatcagcttg catgccggtc gatctagtaa catagatgac accgcgcgcg ataatttatc
     1081 ctagtttgcg cgctatattt tgttttctat cgcgtattaa atgtataatt gcgggactct
     1141 aatcataaaa acccatctca taaataacgt catgcattac atgttaatta ttacatgctt
     1201 aacgtaattc aacagaaatt atatgataat catcgcaaga ccggcaacag gattcaatct
     1261 taagaaactt tattgccaaa tgtttgaacg atctgcttga ctctaggggt catcagattt
     1321 cggtgacggg caggaccgga cggggcggca ccggcaggct gaagtccagc tgccagaaac
     1381 ccacgtcatg ccagttcccg tgcttgaagc cggccgcccg cagcatgccg cggggggcat
     1441 atccgagcgc ctcgtgcatg cgcacgctcg ggtcgttggg cagcccgatg acagcgacca
     1501 cgctcttgaa gccctgtgcc tccagggact tcagcaggtg ggtgtagagc gtggagccca
     1561 gtcccgtccg ctggtggcgg ggggagacgt acacggtcga ctcggccgtc cagtcgtagg
     1621 cgttgcgtgc cttccaggga cccgcgtagg cgatgccggc gacctcgccg tccacctcgg
     1681 cgacgagcca gggatagcgc tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt
     1741 cctgcggctc ggtacggaag ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc
     1801 agaccgccgg catgtccgcc tcggtggcac ggcggatgtc ggccgggcgt cgttctgggc
     1861 tcatggtaga tcccctcgat cgagttgaga gtgaatatga gactctaatt ggataccgag
     1921 gggaatttat ggaacgtcag tggagcattt ttgacaagaa atatttgcta gctgatagtg
     1981 accttaggcg acttttgaac gcgcaataat ggtttctgac gtatgtgctt agctcattaa
     2041 actccagaaa cccgcggctc agtggctcct tcaacgttgc ggttctgtca gttccaaacg
     2101 taaaacggct tgtcccgcgt catcggcggg ggtcataacg tgactccctt aattctccgc
     2161 tcatgatcag attgtcgttt cccgccttcg gtttgggcgc gcccggttca gaagaccaga
     2221 gggctattga gacttttcaa caaagggtaa tatcgggaaa cctcctcgga ttccattgcc
     2281 cagctatctg tcacttcatc gaaaggacag tagaaaagga agatggcttc tacaaatgcc
     2341 atcattgcga taaaggaaag gctatcgttc aagatgcctc taccgacagt ggtcccaaag
     2401 atggaccccc acccacgagg aacatcgtgg aaaaagaaga cgttccaacc acgtcttcaa
     2461 agcaagtgga ttgatgtgat acatggtgga gcacgacact ctcgtctact ccaagaatat
     2521 caaagataca gtctcagaag accagagggc tattgagact tttcaacaaa gggtaatatc
     2581 gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcgaaa ggacagtaga
     2641 aaaggaagat ggcttctaca aatgccatca ttgcgataaa ggaaaggcta tcgttcaaga
     2701 tgcctctacc gacagtggtc ccaaagatgg acccccaccc acgaggaaca tcgtggaaaa
     2761 agaagacgtt ccaaccacgt cttcaaagca agtggattga tgtgatatct ccactgacgt
     2821 aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat aaggaagttc
     2881 atttcatttg gagaggacct cgaggtcgac ggtatcgata agcttataat ggacaagcag
     2941 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     3001 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3061 aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     3121 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3181 aagggagtcc ccatcacaag tttgtacaaa aaagcaggct tcatgtcgac gcgaagacga
     3241 actttactca aagttataat tcttggagac agcggggttg gcaaaacatc gttgatgaat
     3301 caatatgtga ataacaagtt tagtcaacag tacaaagcta cgatcggagc tgattttgtc
     3361 actaaggagc ttcaaattga tgacaggctt gtcacattgc aaatatggga cactgctggg
     3421 ctagagaggt ttcaaagtct tggtgttgct ttctatagag gtgcagattg ttgtgttctt
     3481 gtctatgatg tgaatcactt gaagtcattt gaatctctcg acaattggca caacgagttt
     3541 cttacacggg ctagtccacg tgacccaatg gcattccctt ttatacttct tggtaataag
     3601 gttgatattg atggaggaaa tagccgagtg gtatctgaga agaaggctag agaatggtgt
     3661 gctgaaaagg gaaacatagt ctatttcgag acatcggcta aagaagatta caatgtcgat
     3721 gactccttct tgtgcatcac aaaacttgcc cttgcaaatg aacgcgacca agatatatat
     3781 ttccagccag atactggttc ggtgcctgag caaagaggag gttgtgcttg ctgagaccca
     3841 gctttcttgt acaaagtggt gatgggggtt atccatacga tgttccagat tatgctgtcg
     3901 gcgccggttg gtctcatcct caatttgaaa aataagtcta gagtccgcaa aaatcaccag
     3961 tctctctcta caaatctatc tctctctatt tttctccaga ataatgtgtg agtagttccc
     4021 agataaggga attagggttc ttatagggtt tcgctcatgt gttgagcata taagaaaccc
     4081 ttagtatgta tttgtatttg taaaatactt ctatcaataa aatttctaat tcctaaaacc
     4141 aaaatccagt gaccgggcgg ccgccaccgc ggtggagggg gatcagattg tcgtttcccg
     4201 ccttcagttt aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa
     4261 gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc
     4321 atttgtatgt gtacatcacc gacgagcaag gcaagaccga gcgcctttcc gacgctcacc
     4381 gggctggttg ccctcgccgc tgggctggcg gccgtctatg gccctgcaaa cgcgccagaa
     4441 acgccgtcga agccgtgtgc gagacaccgg ccgccggcgt tgtggatacc tcgcggaaaa
     4501 cttggccctc actgacagat gaggggcgga cgttgacact tgaggggccg actcacccgg
     4561 cgcggcgttg acagatgagg ggcaggctcg atttcggccg gcgacgtgga gctggccagc
     4621 ctcgcaaatc ggcgaaaacg cctgatttta cgcgagtttc ccacagatga tgtggacaag
     4681 cctggggata agtgccctgc ggtattgaca cttgaggggc gcgactactg acagatgagg
     4741 ggcgcgatcc ttgacacttg aggggcagag tgctgacaga tgaggggcgc acctattgac
     4801 atttgagggg ctgtccacag gcagaaaatc cagcatttgc aagggtttcc gcccgttttt
     4861 cggccaccgc taacctgtct tttaacctgc ttttaaacca atatttataa accttgtttt
     4921 taaccagggc tgcgccctgt gcgcgtgacc gcgcacgccg aaggggggtg cccccccttc
     4981 tcgaaccctc ccggcccgct aacgcgggcc tcccatcccc ccaggggctg cgcccctcgg
     5041 ccgcgaacgg cctcacccca aaaatggcag cgctggcagt ccttgccatt gccgggatcg
     5101 gggcagtaac gggatgggcg atcagcccga aagctacccc tatttgttta tttttctaaa
     5161 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5221 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5281 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5341 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5401 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5461 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5521 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5581 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5641 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5701 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5761 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5821 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5881 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5941 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6001 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6061 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6121 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6181 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6241 ccg
//
