LOCUS       sequence-197788-        6196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  split YFP. Plant expression of p35S::ARA6/RabF1-YFPc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6196)
  TITLE     Robatzek Lab plasmids
REFERENCE   2  (bases 1 to 6196)
  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: p35S::ARA6/RabF1-YFPc
FEATURES             Location/Qualifiers
     source          1..6196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      15..603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     504..523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    915..939
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(1033..1285)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1305..1856)
                     /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(1898..2077)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2546..2890
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2834..2854
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2892..2908
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2893..2909
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2925..2949
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3592..3616)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3627..3878
                     /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     3815..3836
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4170..4194
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4306..5015
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5088..5179
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5180..5248,5249..6040)
                     /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(5398..5417)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
       61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      121 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
      181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      661 atggactcta gctagaggat cacaggcagc aacgctctgt catcgttaca atcaacatgc
      721 taccctccgc gagatcatcc gtgtttcaaa cccggcagct tagttgccgt tcttccgaat
      781 agcatcggta acatgagcaa agtctgccgc cttacaacgg ctctcccgct gacgccgtcc
      841 cggactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc
      901 tgttggctgg ctggtggcag gatatattgt ggtgtaaaca aattgacgct tagacaactt
      961 aataacacat tgcggacgtt tttaatgatc gaatactaac gtctctacca gatatcagct
     1021 tgcatgccgg tcgatctagt aacatagatg acaccgcgcg cgataattta tcctagtttg
     1081 cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact ctaatcataa
     1141 aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc ttaacgtaat
     1201 tcaacagaaa ttatatgata atcatcgcaa gaccggcaac aggattcaat cttaagaaac
     1261 tttattgcca aatgtttgaa cgatctgctt gactctaggg gtcatcagat ttcggtgacg
     1321 ggcaggaccg gacggggcgg caccggcagg ctgaagtcca gctgccagaa acccacgtca
     1381 tgccagttcc cgtgcttgaa gccggccgcc cgcagcatgc cgcggggggc atatccgagc
     1441 gcctcgtgca tgcgcacgct cgggtcgttg ggcagcccga tgacagcgac cacgctcttg
     1501 aagccctgtg cctccaggga cttcagcagg tgggtgtaga gcgtggagcc cagtcccgtc
     1561 cgctggtggc ggggggagac gtacacggtc gactcggccg tccagtcgta ggcgttgcgt
     1621 gccttccagg gacccgcgta ggcgatgccg gcgacctcgc cgtccacctc ggcgacgagc
     1681 cagggatagc gctcccgcag acggacgagg tcgtccgtcc actcctgcgg ttcctgcggc
     1741 tcggtacgga agttgaccgt gcttgtctcg atgtagtggt tgacgatggt gcagaccgcc
     1801 ggcatgtccg cctcggtggc acggcggatg tcggccgggc gtcgttctgg gctcatggta
     1861 gatcccctcg atcgagttga gagtgaatat gagactctaa ttggataccg aggggaattt
     1921 atggaacgtc agtggagcat ttttgacaag aaatatttgc tagctgatag tgaccttagg
     1981 cgacttttga acgcgcaata atggtttctg acgtatgtgc ttagctcatt aaactccaga
     2041 aacccgcggc tcagtggctc cttcaacgtt gcggttctgt cagttccaaa cgtaaaacgg
     2101 cttgtcccgc gtcatcggcg ggggtcataa cgtgactccc ttaattctcc gctcatgatc
     2161 agattgtcgt ttcccgcctt cggtttgggc gcgcccggtt cagaagacca gagggctatt
     2221 gagacttttc aacaaagggt aatatcggga aacctcctcg gattccattg cccagctatc
     2281 tgtcacttca tcgaaaggac agtagaaaag gaagatggct tctacaaatg ccatcattgc
     2341 gataaaggaa aggctatcgt tcaagatgcc tctaccgaca gtggtcccaa agatggaccc
     2401 ccacccacga ggaacatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg
     2461 gattgatgtg atacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata
     2521 cagtctcaga agaccagagg gctattgaga cttttcaaca aagggtaata tcgggaaacc
     2581 tcctcggatt ccattgccca gctatctgtc acttcatcga aaggacagta gaaaaggaag
     2641 atggcttcta caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcctcta
     2701 ccgacagtgg tcccaaagat ggacccccac ccacgaggaa catcgtggaa aaagaagacg
     2761 ttccaaccac gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg
     2821 acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt
     2881 tggagaggac ctcgaggtcg acggtatcga taagcttccc atcaacaagt ttgtacaaaa
     2941 aagcaggctc cgcggccgcc cccttcacca tgggatgtgc ttcttctctt ccagatagga
     3001 actctggaac tttaagtggt cttagcaatt cagagaatgc tgttccagct gatgccaaaa
     3061 atctacgtgt gaagttagtt ctattaggag actctggtgt tggtaaaagt tgtattgtcc
     3121 ttcgatttgt acgtggtcag tttgacgcta catctaaggt aactgttgga gcctcgttct
     3181 tgtcccaaac tatagcattg caagactcta ccacagtgaa gtttgaaata tgggatacag
     3241 caggacagga gaggtattct gctcttgcac cactatacta ccgtggagct ggagttgctg
     3301 ttattgtgta tgatataaca agccctgaat cgttcaagaa agcacagtat tgggttaagg
     3361 aactgcaaaa gcatggaagc ccagatattg ttatggctct ggttggtaac aaagctgatc
     3421 tacatgaaaa aagagaagta cctactgagg atggaatgga gcttgcagag aagaacggca
     3481 tgttcttcat tgagacgtca gccaagacag ccgataacat aaatcaactg tttgaggaaa
     3541 ttggcaagag gctacctcgt cctgctcctt cgtcaaaggg tgggcgcgcc gacccagctt
     3601 tcttgtacaa agtggttgat gggatggaca agcagaagaa cggcatcaag gccaacttca
     3661 agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca
     3721 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg
     3781 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     3841 ccgccgggat cactctcggc atggacgagc tgtacaagta atctagagtc cgcaaaaatc
     3901 accagtctct ctctacaaat ctatctctct ctatttttct ccagaataat gtgtgagtag
     3961 ttcccagata agggaattag ggttcttata gggtttcgct catgtgttga gcatataaga
     4021 aacccttagt atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta
     4081 aaaccaaaat ccagtgaccg ggcggccgcc accgcggtgg agggggatca gattgtcgtt
     4141 tcccgccttc agtttaaact atcagtgttt gacaggatat attggcgggt aaacctaaga
     4201 gaaaagagcg tttattagaa taatcggata tttaaaaggg cgtgaaaagg tttatccgtt
     4261 cgtccatttg tatgtgtaca tcaccgacga gcaaggcaag accgagcgcc tttccgacgc
     4321 tcaccgggct ggttgccctc gccgctgggc tggcggccgt ctatggccct gcaaacgcgc
     4381 cagaaacgcc gtcgaagccg tgtgcgagac accggccgcc ggcgttgtgg atacctcgcg
     4441 gaaaacttgg ccctcactga cagatgaggg gcggacgttg acacttgagg ggccgactca
     4501 cccggcgcgg cgttgacaga tgaggggcag gctcgatttc ggccggcgac gtggagctgg
     4561 ccagcctcgc aaatcggcga aaacgcctga ttttacgcga gtttcccaca gatgatgtgg
     4621 acaagcctgg ggataagtgc cctgcggtat tgacacttga ggggcgcgac tactgacaga
     4681 tgaggggcgc gatccttgac acttgagggg cagagtgctg acagatgagg ggcgcaccta
     4741 ttgacatttg aggggctgtc cacaggcaga aaatccagca tttgcaaggg tttccgcccg
     4801 tttttcggcc accgctaacc tgtcttttaa cctgctttta aaccaatatt tataaacctt
     4861 gtttttaacc agggctgcgc cctgtgcgcg tgaccgcgca cgccgaaggg gggtgccccc
     4921 ccttctcgaa ccctcccggc ccgctaacgc gggcctccca tccccccagg ggctgcgccc
     4981 ctcggccgcg aacggcctca ccccaaaaat ggcagcgctg gcagtccttg ccattgccgg
     5041 gatcggggca gtaacgggat gggcgatcag cccgaaagct acccctattt gtttattttt
     5101 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5161 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5221 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5281 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5341 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5401 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5461 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5521 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5581 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5641 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5701 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5761 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5821 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5881 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5941 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6001 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6061 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6121 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6181 agaccccgta gaaaag
//
