LOCUS       sequence-196854-        6166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  split YFP. Plant expression of CBL5-YFPc-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6166)
  TITLE     Robatzek Lab plasmids
REFERENCE   2  (bases 1 to 6166)
  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-CBL5-YFPc-1
FEATURES             Location/Qualifiers
     source          1..6166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      5..593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     494..513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    905..929
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(1023..1275)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1295..1846)
                     /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(1888..2067)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2536..2880
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2824..2844
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2882..2898
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2883..2899
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2915..2939
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3552..3576)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3587..3838
                     /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     3775..3796
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4130..4154
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4266..4975
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5048..5139
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5140..5208,5209..6000)
                     /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(5358..5377)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
       61 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      121 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
      181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atggactcta
      661 gctagaggat cacaggcagc aacgctctgt catcgttaca atcaacatgc taccctccgc
      721 gagatcatcc gtgtttcaaa cccggcagct tagttgccgt tcttccgaat agcatcggta
      781 acatgagcaa agtctgccgc cttacaacgg ctctcccgct gacgccgtcc cggactgatg
      841 ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc tgttggctgg
      901 ctggtggcag gatatattgt ggtgtaaaca aattgacgct tagacaactt aataacacat
      961 tgcggacgtt tttaatgatc gaatactaac gtctctacca gatatcagct tgcatgccgg
     1021 tcgatctagt aacatagatg acaccgcgcg cgataattta tcctagtttg cgcgctatat
     1081 tttgttttct atcgcgtatt aaatgtataa ttgcgggact ctaatcataa aaacccatct
     1141 cataaataac gtcatgcatt acatgttaat tattacatgc ttaacgtaat tcaacagaaa
     1201 ttatatgata atcatcgcaa gaccggcaac aggattcaat cttaagaaac tttattgcca
     1261 aatgtttgaa cgatctgctt gactctaggg gtcatcagat ttcggtgacg ggcaggaccg
     1321 gacggggcgg caccggcagg ctgaagtcca gctgccagaa acccacgtca tgccagttcc
     1381 cgtgcttgaa gccggccgcc cgcagcatgc cgcggggggc atatccgagc gcctcgtgca
     1441 tgcgcacgct cgggtcgttg ggcagcccga tgacagcgac cacgctcttg aagccctgtg
     1501 cctccaggga cttcagcagg tgggtgtaga gcgtggagcc cagtcccgtc cgctggtggc
     1561 ggggggagac gtacacggtc gactcggccg tccagtcgta ggcgttgcgt gccttccagg
     1621 gacccgcgta ggcgatgccg gcgacctcgc cgtccacctc ggcgacgagc cagggatagc
     1681 gctcccgcag acggacgagg tcgtccgtcc actcctgcgg ttcctgcggc tcggtacgga
     1741 agttgaccgt gcttgtctcg atgtagtggt tgacgatggt gcagaccgcc ggcatgtccg
     1801 cctcggtggc acggcggatg tcggccgggc gtcgttctgg gctcatggta gatcccctcg
     1861 atcgagttga gagtgaatat gagactctaa ttggataccg aggggaattt atggaacgtc
     1921 agtggagcat ttttgacaag aaatatttgc tagctgatag tgaccttagg cgacttttga
     1981 acgcgcaata atggtttctg acgtatgtgc ttagctcatt aaactccaga aacccgcggc
     2041 tcagtggctc cttcaacgtt gcggttctgt cagttccaaa cgtaaaacgg cttgtcccgc
     2101 gtcatcggcg ggggtcataa cgtgactccc ttaattctcc gctcatgatc agattgtcgt
     2161 ttcccgcctt cggtttgggc gcgcccggtt cagaagacca gagggctatt gagacttttc
     2221 aacaaagggt aatatcggga aacctcctcg gattccattg cccagctatc tgtcacttca
     2281 tcgaaaggac agtagaaaag gaagatggct tctacaaatg ccatcattgc gataaaggaa
     2341 aggctatcgt tcaagatgcc tctaccgaca gtggtcccaa agatggaccc ccacccacga
     2401 ggaacatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg
     2461 atacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata cagtctcaga
     2521 agaccagagg gctattgaga cttttcaaca aagggtaata tcgggaaacc tcctcggatt
     2581 ccattgccca gctatctgtc acttcatcga aaggacagta gaaaaggaag atggcttcta
     2641 caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcctcta ccgacagtgg
     2701 tcccaaagat ggacccccac ccacgaggaa catcgtggaa aaagaagacg ttccaaccac
     2761 gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc
     2821 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac
     2881 ctcgaggtcg acggtatcga taagcttccc atcaacaagt ttgtacaaaa aagcaggctt
     2941 gatgggatgt gtttgcagca agcaattaga aggaagaagg caagaagaca tttcgcttct
     3001 tgcttctcaa acctttttta gcgaggctga ggttgaagtt ttgcatggat tgttcatcaa
     3061 actcacttct tgtcttagca acgacaatct tttgaccaaa gaaaaatttc aatttatctt
     3121 gatcaagaat accaaaaagc gcagtctttc tgcagaacgg atttttggat tattcgatat
     3181 gagaaacgat ggggctatag attttggaga gtttgtacac acactcaaca tttttcaccc
     3241 aaactcatca ccaagagata aggccatatt tgcatttaga ttatacgata ctcgtgagac
     3301 tggattcatt gaaccggagg aggtgaaaga gatgataata gacgttctag aggaatcaga
     3361 acttatgcta tccgagagta tcattgattc tatcgtgtct aagacatttg aagaggccga
     3421 ttggaaaaaa gatgggataa tagatttgga agaatgggag aattttgttg ccacctatcc
     3481 tttgacacta aagaacatga ctatcccttt cttgaaggac attccgagga ttttcccaac
     3541 ctttctccgg tacccagctt tcttgtacaa agtggttgat gggatggaca agcagaagaa
     3601 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
     3661 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     3721 ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     3781 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     3841 atctagagtc cgcaaaaatc accagtctct ctctacaaat ctatctctct ctatttttct
     3901 ccagaataat gtgtgagtag ttcccagata agggaattag ggttcttata gggtttcgct
     3961 catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc
     4021 aataaaattt ctaattccta aaaccaaaat ccagtgaccg ggcggccgcc accgcggtgg
     4081 agggggatca gattgtcgtt tcccgccttc agtttaaact atcagtgttt gacaggatat
     4141 attggcgggt aaacctaaga gaaaagagcg tttattagaa taatcggata tttaaaaggg
     4201 cgtgaaaagg tttatccgtt cgtccatttg tatgtgtaca tcaccgacga gcaaggcaag
     4261 accgagcgcc tttccgacgc tcaccgggct ggttgccctc gccgctgggc tggcggccgt
     4321 ctatggccct gcaaacgcgc cagaaacgcc gtcgaagccg tgtgcgagac accggccgcc
     4381 ggcgttgtgg atacctcgcg gaaaacttgg ccctcactga cagatgaggg gcggacgttg
     4441 acacttgagg ggccgactca cccggcgcgg cgttgacaga tgaggggcag gctcgatttc
     4501 ggccggcgac gtggagctgg ccagcctcgc aaatcggcga aaacgcctga ttttacgcga
     4561 gtttcccaca gatgatgtgg acaagcctgg ggataagtgc cctgcggtat tgacacttga
     4621 ggggcgcgac tactgacaga tgaggggcgc gatccttgac acttgagggg cagagtgctg
     4681 acagatgagg ggcgcaccta ttgacatttg aggggctgtc cacaggcaga aaatccagca
     4741 tttgcaaggg tttccgcccg tttttcggcc accgctaacc tgtcttttaa cctgctttta
     4801 aaccaatatt tataaacctt gtttttaacc agggctgcgc cctgtgcgcg tgaccgcgca
     4861 cgccgaaggg gggtgccccc ccttctcgaa ccctcccggc ccgctaacgc gggcctccca
     4921 tccccccagg ggctgcgccc ctcggccgcg aacggcctca ccccaaaaat ggcagcgctg
     4981 gcagtccttg ccattgccgg gatcggggca gtaacgggat gggcgatcag cccgaaagct
     5041 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5101 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5161 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5221 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5281 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5341 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5401 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5461 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5521 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5581 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5641 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5701 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5761 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5821 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5881 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5941 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6001 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6061 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6121 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggat
//
