LOCUS       sequence-197600-        7408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  split YFP. Plant expression of CERK1 D441V-YFPc-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7408)
  TITLE     Robatzek Lab plasmids
REFERENCE   2  (bases 1 to 7408)
  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-CERK1 D441V-YFPc-1
FEATURES             Location/Qualifiers
     source          1..7408
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      9..597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     498..517
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    909..933
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(1027..1279)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1299..1850)
                     /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(1892..2071)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2540..2884
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2828..2848
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2886..2902
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2887..2903
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2919..2943
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4798..4822)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4833..5084
                     /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     5021..5042
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5376..5400
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      5512..6221
                     /label=oriV
                     /note="incP origin of replication"
     promoter        6294..6385
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6386..6454,6455..7246)
                     /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(6604..6623)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
       61 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      121 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      181 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      241 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      301 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      361 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      421 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      481 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      541 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      601 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggac
      661 tctagctaga ggatcacagg cagcaacgct ctgtcatcgt tacaatcaac atgctaccct
      721 ccgcgagatc atccgtgttt caaacccggc agcttagttg ccgttcttcc gaatagcatc
      781 ggtaacatga gcaaagtctg ccgccttaca acggctctcc cgctgacgcc gtcccggact
      841 gatgggctgc ctgtatcgag tggtgatttt gtgccgagct gccggtcggg gagctgttgg
      901 ctggctggtg gcaggatata ttgtggtgta aacaaattga cgcttagaca acttaataac
      961 acattgcgga cgtttttaat gatcgaatac taacgtctct accagatatc agcttgcatg
     1021 ccggtcgatc tagtaacata gatgacaccg cgcgcgataa tttatcctag tttgcgcgct
     1081 atattttgtt ttctatcgcg tattaaatgt ataattgcgg gactctaatc ataaaaaccc
     1141 atctcataaa taacgtcatg cattacatgt taattattac atgcttaacg taattcaaca
     1201 gaaattatat gataatcatc gcaagaccgg caacaggatt caatcttaag aaactttatt
     1261 gccaaatgtt tgaacgatct gcttgactct aggggtcatc agatttcggt gacgggcagg
     1321 accggacggg gcggcaccgg caggctgaag tccagctgcc agaaacccac gtcatgccag
     1381 ttcccgtgct tgaagccggc cgcccgcagc atgccgcggg gggcatatcc gagcgcctcg
     1441 tgcatgcgca cgctcgggtc gttgggcagc ccgatgacag cgaccacgct cttgaagccc
     1501 tgtgcctcca gggacttcag caggtgggtg tagagcgtgg agcccagtcc cgtccgctgg
     1561 tggcgggggg agacgtacac ggtcgactcg gccgtccagt cgtaggcgtt gcgtgccttc
     1621 cagggacccg cgtaggcgat gccggcgacc tcgccgtcca cctcggcgac gagccaggga
     1681 tagcgctccc gcagacggac gaggtcgtcc gtccactcct gcggttcctg cggctcggta
     1741 cggaagttga ccgtgcttgt ctcgatgtag tggttgacga tggtgcagac cgccggcatg
     1801 tccgcctcgg tggcacggcg gatgtcggcc gggcgtcgtt ctgggctcat ggtagatccc
     1861 ctcgatcgag ttgagagtga atatgagact ctaattggat accgagggga atttatggaa
     1921 cgtcagtgga gcatttttga caagaaatat ttgctagctg atagtgacct taggcgactt
     1981 ttgaacgcgc aataatggtt tctgacgtat gtgcttagct cattaaactc cagaaacccg
     2041 cggctcagtg gctccttcaa cgttgcggtt ctgtcagttc caaacgtaaa acggcttgtc
     2101 ccgcgtcatc ggcgggggtc ataacgtgac tcccttaatt ctccgctcat gatcagattg
     2161 tcgtttcccg ccttcggttt gggcgcgccc ggttcagaag accagagggc tattgagact
     2221 tttcaacaaa gggtaatatc gggaaacctc ctcggattcc attgcccagc tatctgtcac
     2281 ttcatcgaaa ggacagtaga aaaggaagat ggcttctaca aatgccatca ttgcgataaa
     2341 ggaaaggcta tcgttcaaga tgcctctacc gacagtggtc ccaaagatgg acccccaccc
     2401 acgaggaaca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca agtggattga
     2461 tgtgatacat ggtggagcac gacactctcg tctactccaa gaatatcaaa gatacagtct
     2521 cagaagacca gagggctatt gagacttttc aacaaagggt aatatcggga aacctcctcg
     2581 gattccattg cccagctatc tgtcacttca tcgaaaggac agtagaaaag gaagatggct
     2641 tctacaaatg ccatcattgc gataaaggaa aggctatcgt tcaagatgcc tctaccgaca
     2701 gtggtcccaa agatggaccc ccacccacga ggaacatcgt ggaaaaagaa gacgttccaa
     2761 ccacgtcttc aaagcaagtg gattgatgtg atatctccac tgacgtaagg gatgacgcac
     2821 aatcccacta tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga
     2881 ggacctcgag gtcgacggta tcgataagct tcccatcaac aagtttgtac aaaaaagcag
     2941 gcttgatgaa gctaaagatt tctctaatcg ctccgattct tcttctcttc tcattcttct
     3001 tcgccgtgga atctaagtgc aggactagct gtcctttagc tctagcttcg tactatctcg
     3061 agaacggaac aacactctcc gtcatcaacc aaaacctcaa ttcttcaatc gcgccttacg
     3121 atcaaatcaa tttcgatcca atcctcaggt acaacagtaa cattaaagac aaagatagaa
     3181 tccagatggg ctctagggtt cttgtacctt tcccttgcga atgtcaacct ggtgatttcc
     3241 tagggcacaa tttcagctac agtgttcgac aggaagatac ttacgaaaga gtcgcgatta
     3301 gtaattacgc gaatctcacg acgatggagt cgttacaggc gaggaatcct tttccggcga
     3361 ctaacatacc tctctctgcg acgcttaatg tattggtgaa ttgttcttgt ggtgatgaga
     3421 gtgtttcgaa agattttggt ttgtttgtta cgtatccgct tcgtcctgaa gacagtctca
     3481 gttctattgc gagatcttcc ggtgtatcgg cggatattct gcagagatat aatcccggtg
     3541 ttaattttaa ctccgggaat ggaatcgttt atgtgcctgg aagagatcca aatggtgcat
     3601 ttccaccatt caaatcaagt aaacaagatg gtgttggtgc tggagttatt gctggtatag
     3661 ttataggagt gattgtggct ttgttgttga tcttgtttat cgtatattat gcttaccgga
     3721 agaataagtc gaagggtgat tcgttttctt cttctattcc gttgtctact aaggctgatc
     3781 atgcttcttc tactagtctc caaagtggag gtttgggtgg tgccggagtg tctcctggca
     3841 ttgctgccat aagcgtggac aaatctgttg agttttcgtt ggaggaacta gcaaaggcta
     3901 ctgataattt caatttgtct tttaagattg ggcaaggtgg ttttggggct gtttactatg
     3961 cagagctgag aggagaaaaa gctgcgatta agaagatgga catggaggca tcgaaacagt
     4021 tcttggcgga actaaaagtc ttaacgcgtg tacatcatgt caacctggtt cgcctgattg
     4081 gatattgtgt tgagggatca cttttcttgg tgtatgaata tgttgagaat ggtaaccttg
     4141 gacaacattt acatgggtca ggacgagaac cattaccgtg gactaagaga gtgcagattg
     4201 cactagactc agctagaggt ttagaatata tccacgagca cacggttcca gtttatgtcc
     4261 atagggtcat taaatctgcc aatattttga tagaccagaa attccgagca aaggtagcag
     4321 atttcgggtt aacaaaactg acagaagttg gaggttcagc aactcggggt gcaatgggta
     4381 catttggtta catggcacca gagactgttt atggagaagt gtctgcaaaa gtagatgtat
     4441 atgcatttgg agttgtcctt tacgaattga tttctgcgaa aggtgcggtt gtcaaaatga
     4501 cagaagccgt tggtgaattt agaggccttg ttggtgtgtt cgaagaatca ttcaaggaaa
     4561 ccgacaaaga agaagcacta cgcaagatta tagacccgag gctcggtgat agttacccgt
     4621 ttgattcggt atacaagatg gcggaattag ggaaagcatg tacacaagag aatgcgcagc
     4681 tacgtccgag tatgagatac attgtggttg ctttatccac tctcttttcg tctaccggaa
     4741 attgggatgt tggaaacttc caaaacgaag atttagtcag tcttatgtcc ggccggtacc
     4801 cagctttctt gtacaaagtg gttgatggga tggacaagca gaagaacggc atcaaggcca
     4861 acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc
     4921 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     4981 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     5041 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatct agagtccgca
     5101 aaaatcacca gtctctctct acaaatctat ctctctctat ttttctccag aataatgtgt
     5161 gagtagttcc cagataaggg aattagggtt cttatagggt ttcgctcatg tgttgagcat
     5221 ataagaaacc cttagtatgt atttgtattt gtaaaatact tctatcaata aaatttctaa
     5281 ttcctaaaac caaaatccag tgaccgggcg gccgccaccg cggtggaggg ggatcagatt
     5341 gtcgtttccc gccttcagtt taaactatca gtgtttgaca ggatatattg gcgggtaaac
     5401 ctaagagaaa agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta
     5461 tccgttcgtc catttgtatg tgtacatcac cgacgagcaa ggcaagaccg agcgcctttc
     5521 cgacgctcac cgggctggtt gccctcgccg ctgggctggc ggccgtctat ggccctgcaa
     5581 acgcgccaga aacgccgtcg aagccgtgtg cgagacaccg gccgccggcg ttgtggatac
     5641 ctcgcggaaa acttggccct cactgacaga tgaggggcgg acgttgacac ttgaggggcc
     5701 gactcacccg gcgcggcgtt gacagatgag gggcaggctc gatttcggcc ggcgacgtgg
     5761 agctggccag cctcgcaaat cggcgaaaac gcctgatttt acgcgagttt cccacagatg
     5821 atgtggacaa gcctggggat aagtgccctg cggtattgac acttgagggg cgcgactact
     5881 gacagatgag gggcgcgatc cttgacactt gaggggcaga gtgctgacag atgaggggcg
     5941 cacctattga catttgaggg gctgtccaca ggcagaaaat ccagcatttg caagggtttc
     6001 cgcccgtttt tcggccaccg ctaacctgtc ttttaacctg cttttaaacc aatatttata
     6061 aaccttgttt ttaaccaggg ctgcgccctg tgcgcgtgac cgcgcacgcc gaaggggggt
     6121 gccccccctt ctcgaaccct cccggcccgc taacgcgggc ctcccatccc cccaggggct
     6181 gcgcccctcg gccgcgaacg gcctcacccc aaaaatggca gcgctggcag tccttgccat
     6241 tgccgggatc ggggcagtaa cgggatgggc gatcagcccg aaagctaccc ctatttgttt
     6301 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6361 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6421 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6481 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6541 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6601 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6661 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6721 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6781 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6841 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6901 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6961 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     7021 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     7081 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     7141 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     7201 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     7261 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     7321 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7381 agcgtcagac cccgtagaaa agatcaaa
//
