LOCUS       sequence-196616-        6850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  split YFP. Plant expression of Snrk2.3-YFPn-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6850)
  TITLE     Robatzek Lab plasmids
REFERENCE   2  (bases 1 to 6850)
  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: pAM-35s-Snrk2.3-YFPn-1
FEATURES             Location/Qualifiers
     source          1..6850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      18..606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     507..526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    918..942
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(1036..1288)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1308..1859)
                     /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(1901..2080)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2549..2893
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2837..2857
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2895..2911
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2896..2912
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2928..2952
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4039..4063)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(4071..4073,4074..4076,4077..4535)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMA"
     primer_bind     complement(4116..4137)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4377..4396)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4827..4851
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4963..5672
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5745..5836
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5837..5905,5906..6697)
                     /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(6055..6074)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
       61 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      121 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
      181 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      241 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      301 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      361 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      421 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      481 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      541 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      601 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      661 cacatggact ctagctagag gatcacaggc agcaacgctc tgtcatcgtt acaatcaaca
      721 tgctaccctc cgcgagatca tccgtgtttc aaacccggca gcttagttgc cgttcttccg
      781 aatagcatcg gtaacatgag caaagtctgc cgccttacaa cggctctccc gctgacgccg
      841 tcccggactg atgggctgcc tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg
      901 agctgttggc tggctggtgg caggatatat tgtggtgtaa acaaattgac gcttagacaa
      961 cttaataaca cattgcggac gtttttaatg atcgaatact aacgtctcta ccagatatca
     1021 gcttgcatgc cggtcgatct agtaacatag atgacaccgc gcgcgataat ttatcctagt
     1081 ttgcgcgcta tattttgttt tctatcgcgt attaaatgta taattgcggg actctaatca
     1141 taaaaaccca tctcataaat aacgtcatgc attacatgtt aattattaca tgcttaacgt
     1201 aattcaacag aaattatatg ataatcatcg caagaccggc aacaggattc aatcttaaga
     1261 aactttattg ccaaatgttt gaacgatctg cttgactcta ggggtcatca gatttcggtg
     1321 acgggcagga ccggacgggg cggcaccggc aggctgaagt ccagctgcca gaaacccacg
     1381 tcatgccagt tcccgtgctt gaagccggcc gcccgcagca tgccgcgggg ggcatatccg
     1441 agcgcctcgt gcatgcgcac gctcgggtcg ttgggcagcc cgatgacagc gaccacgctc
     1501 ttgaagccct gtgcctccag ggacttcagc aggtgggtgt agagcgtgga gcccagtccc
     1561 gtccgctggt ggcgggggga gacgtacacg gtcgactcgg ccgtccagtc gtaggcgttg
     1621 cgtgccttcc agggacccgc gtaggcgatg ccggcgacct cgccgtccac ctcggcgacg
     1681 agccagggat agcgctcccg cagacggacg aggtcgtccg tccactcctg cggttcctgc
     1741 ggctcggtac ggaagttgac cgtgcttgtc tcgatgtagt ggttgacgat ggtgcagacc
     1801 gccggcatgt ccgcctcggt ggcacggcgg atgtcggccg ggcgtcgttc tgggctcatg
     1861 gtagatcccc tcgatcgagt tgagagtgaa tatgagactc taattggata ccgaggggaa
     1921 tttatggaac gtcagtggag catttttgac aagaaatatt tgctagctga tagtgacctt
     1981 aggcgacttt tgaacgcgca ataatggttt ctgacgtatg tgcttagctc attaaactcc
     2041 agaaacccgc ggctcagtgg ctccttcaac gttgcggttc tgtcagttcc aaacgtaaaa
     2101 cggcttgtcc cgcgtcatcg gcgggggtca taacgtgact cccttaattc tccgctcatg
     2161 atcagattgt cgtttcccgc cttcggtttg ggcgcgcccg gttcagaaga ccagagggct
     2221 attgagactt ttcaacaaag ggtaatatcg ggaaacctcc tcggattcca ttgcccagct
     2281 atctgtcact tcatcgaaag gacagtagaa aaggaagatg gcttctacaa atgccatcat
     2341 tgcgataaag gaaaggctat cgttcaagat gcctctaccg acagtggtcc caaagatgga
     2401 cccccaccca cgaggaacat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa
     2461 gtggattgat gtgatacatg gtggagcacg acactctcgt ctactccaag aatatcaaag
     2521 atacagtctc agaagaccag agggctattg agacttttca acaaagggta atatcgggaa
     2581 acctcctcgg attccattgc ccagctatct gtcacttcat cgaaaggaca gtagaaaagg
     2641 aagatggctt ctacaaatgc catcattgcg ataaaggaaa ggctatcgtt caagatgcct
     2701 ctaccgacag tggtcccaaa gatggacccc cacccacgag gaacatcgtg gaaaaagaag
     2761 acgttccaac cacgtcttca aagcaagtgg attgatgtga tatctccact gacgtaaggg
     2821 atgacgcaca atcccactat ccttcgcaag acccttcctc tatataagga agttcatttc
     2881 atttggagag gacctcgagg tcgacggtat cgataagctt cccatcaaca agtttgtaca
     2941 aaaaagcagg cttgatggat cgagctccgg tgaccacagg accgttggat atgccgatta
     3001 tgcacgacag tgatcgatat gacttcgtta aggatattgg ttctggtaat ttcggtgttg
     3061 ctcgtcttat gagagataaa ctcactaaag agcttgttgc tgtcaagtac atcgagagag
     3121 gagacaagat tgatgaaaat gttcaaaggg agatcattaa ccacaggtca ctaaggcatc
     3181 ctaatattgt cagatttaaa gaggtcattt tgacgccgac tcatctggct atcataatgg
     3241 aatatgcttc tggcggtgaa ctttacgagc ggatttgcaa tgcaggacgg tttagtgaag
     3301 atgaggctcg gttcttcttt cagcagcttc tatctggagt cagttattgt cattcgatgc
     3361 aaatttgcca tcgtgacctg aagctagaga atacattgtt ggatggaagt cctgctcctc
     3421 gattaaaaat ttgtgatttt ggatattcaa agtcttctgt tcttcattca caaccaaagt
     3481 caactgttgg tactcctgca tacatcgctc cagaggtact gcttcgtcag gaatatgatg
     3541 gcaagattgc agatgtatgg tcatgtggtg tgaccttata cgtcatgttg gttggagcgt
     3601 atccgttcga agatccagaa gagccaagag actatcggaa aacaatacag agaatcctta
     3661 gcgttaaata ctcaatccct gatgacatac ggatatcacc tgaatgctgt catcttattt
     3721 caagaatctt cgtggctgat cccgctacca gaataagcat accagagatc aaaacccata
     3781 gttggttctt gaagaatctc cctgctgatc taatgaacga gagcaacaca ggaagccagt
     3841 tccaggagcc tgaacaacca atgcaaagcc ttgacacaat catgcaaatc atctctgaag
     3901 ccacaattcc cgctgttcga aaccgttgcc tagacgattt catgactgac aatcttgatc
     3961 ttgacgatga catggatgac tttgactctg aatctgaaat cgacattgac agtagcggag
     4021 agatagttta cgctctctac ccagctttct tgtacaaagt ggttgatggg atggtgagca
     4081 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     4141 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     4201 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     4261 ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     4321 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     4381 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     4441 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     4501 acaactacaa cagccacaac gtctatatca tggcctaatc tagagtccgc aaaaatcacc
     4561 agtctctctc tacaaatcta tctctctcta tttttctcca gaataatgtg tgagtagttc
     4621 ccagataagg gaattagggt tcttataggg tttcgctcat gtgttgagca tataagaaac
     4681 ccttagtatg tatttgtatt tgtaaaatac ttctatcaat aaaatttcta attcctaaaa
     4741 ccaaaatcca gtgaccgggc ggccgccacc gcggtggagg gggatcagat tgtcgtttcc
     4801 cgccttcagt ttaaactatc agtgtttgac aggatatatt ggcgggtaaa cctaagagaa
     4861 aagagcgttt attagaataa tcggatattt aaaagggcgt gaaaaggttt atccgttcgt
     4921 ccatttgtat gtgtacatca ccgacgagca aggcaagacc gagcgccttt ccgacgctca
     4981 ccgggctggt tgccctcgcc gctgggctgg cggccgtcta tggccctgca aacgcgccag
     5041 aaacgccgtc gaagccgtgt gcgagacacc ggccgccggc gttgtggata cctcgcggaa
     5101 aacttggccc tcactgacag atgaggggcg gacgttgaca cttgaggggc cgactcaccc
     5161 ggcgcggcgt tgacagatga ggggcaggct cgatttcggc cggcgacgtg gagctggcca
     5221 gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt tcccacagat gatgtggaca
     5281 agcctgggga taagtgccct gcggtattga cacttgaggg gcgcgactac tgacagatga
     5341 ggggcgcgat ccttgacact tgaggggcag agtgctgaca gatgaggggc gcacctattg
     5401 acatttgagg ggctgtccac aggcagaaaa tccagcattt gcaagggttt ccgcccgttt
     5461 ttcggccacc gctaacctgt cttttaacct gcttttaaac caatatttat aaaccttgtt
     5521 tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc cgaagggggg tgccccccct
     5581 tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc ccccaggggc tgcgcccctc
     5641 ggccgcgaac ggcctcaccc caaaaatggc agcgctggca gtccttgcca ttgccgggat
     5701 cggggcagta acgggatggg cgatcagccc gaaagctacc cctatttgtt tatttttcta
     5761 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5821 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5881 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5941 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     6001 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     6061 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     6121 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     6181 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6241 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6301 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6361 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6421 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6481 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6541 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6601 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6661 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6721 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6781 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6841 ccccgtagaa
//
