LOCUS       sequence-196620-        6196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  split YFP. Plant expression of CBL1-YFPc-1.
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: pAM-35s-CBL1-YFPc-1
FEATURES             Location/Qualifiers
     source          1..6196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      33..621
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     522..541
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    933..957
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(1051..1303)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1323..1874)
                     /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(1916..2095)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2564..2908
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2852..2872
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2910..2926
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2911..2927
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2943..2967
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3610..3634)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3645..3896
                     /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     3833..3854
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4188..4212
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4324..5033
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5106..5197
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5198..5266,5267..6058)
                     /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(5416..5435)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
       61 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
      121 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
      181 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
      241 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
      301 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
      361 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
      421 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
      481 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
      541 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
      601 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
      661 tgctggcctt ttgctcacat ggactctagc tagaggatca caggcagcaa cgctctgtca
      721 tcgttacaat caacatgcta ccctccgcga gatcatccgt gtttcaaacc cggcagctta
      781 gttgccgttc ttccgaatag catcggtaac atgagcaaag tctgccgcct tacaacggct
      841 ctcccgctga cgccgtcccg gactgatggg ctgcctgtat cgagtggtga ttttgtgccg
      901 agctgccggt cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacaaa
      961 ttgacgctta gacaacttaa taacacattg cggacgtttt taatgatcga atactaacgt
     1021 ctctaccaga tatcagcttg catgccggtc gatctagtaa catagatgac accgcgcgcg
     1081 ataatttatc ctagtttgcg cgctatattt tgttttctat cgcgtattaa atgtataatt
     1141 gcgggactct aatcataaaa acccatctca taaataacgt catgcattac atgttaatta
     1201 ttacatgctt aacgtaattc aacagaaatt atatgataat catcgcaaga ccggcaacag
     1261 gattcaatct taagaaactt tattgccaaa tgtttgaacg atctgcttga ctctaggggt
     1321 catcagattt cggtgacggg caggaccgga cggggcggca ccggcaggct gaagtccagc
     1381 tgccagaaac ccacgtcatg ccagttcccg tgcttgaagc cggccgcccg cagcatgccg
     1441 cggggggcat atccgagcgc ctcgtgcatg cgcacgctcg ggtcgttggg cagcccgatg
     1501 acagcgacca cgctcttgaa gccctgtgcc tccagggact tcagcaggtg ggtgtagagc
     1561 gtggagccca gtcccgtccg ctggtggcgg ggggagacgt acacggtcga ctcggccgtc
     1621 cagtcgtagg cgttgcgtgc cttccaggga cccgcgtagg cgatgccggc gacctcgccg
     1681 tccacctcgg cgacgagcca gggatagcgc tcccgcagac ggacgaggtc gtccgtccac
     1741 tcctgcggtt cctgcggctc ggtacggaag ttgaccgtgc ttgtctcgat gtagtggttg
     1801 acgatggtgc agaccgccgg catgtccgcc tcggtggcac ggcggatgtc ggccgggcgt
     1861 cgttctgggc tcatggtaga tcccctcgat cgagttgaga gtgaatatga gactctaatt
     1921 ggataccgag gggaatttat ggaacgtcag tggagcattt ttgacaagaa atatttgcta
     1981 gctgatagtg accttaggcg acttttgaac gcgcaataat ggtttctgac gtatgtgctt
     2041 agctcattaa actccagaaa cccgcggctc agtggctcct tcaacgttgc ggttctgtca
     2101 gttccaaacg taaaacggct tgtcccgcgt catcggcggg ggtcataacg tgactccctt
     2161 aattctccgc tcatgatcag attgtcgttt cccgccttcg gtttgggcgc gcccggttca
     2221 gaagaccaga gggctattga gacttttcaa caaagggtaa tatcgggaaa cctcctcgga
     2281 ttccattgcc cagctatctg tcacttcatc gaaaggacag tagaaaagga agatggcttc
     2341 tacaaatgcc atcattgcga taaaggaaag gctatcgttc aagatgcctc taccgacagt
     2401 ggtcccaaag atggaccccc acccacgagg aacatcgtgg aaaaagaaga cgttccaacc
     2461 acgtcttcaa agcaagtgga ttgatgtgat acatggtgga gcacgacact ctcgtctact
     2521 ccaagaatat caaagataca gtctcagaag accagagggc tattgagact tttcaacaaa
     2581 gggtaatatc gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcgaaa
     2641 ggacagtaga aaaggaagat ggcttctaca aatgccatca ttgcgataaa ggaaaggcta
     2701 tcgttcaaga tgcctctacc gacagtggtc ccaaagatgg acccccaccc acgaggaaca
     2761 tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca agtggattga tgtgatatct
     2821 ccactgacgt aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat
     2881 aaggaagttc atttcatttg gagaggacct cgaggtcgac ggtatcgata agcttcccat
     2941 caacaagttt gtacaaaaaa gcaggcttga tgggctgctt ccactcaaag gcagcaaaag
     3001 aatttcgagg acacgaagac cctgttaagc ttgcctctga gacagctttt agtgtgagtg
     3061 aggttgaagc attatttgag ctattcaaga gcataagcag ttcggttgtt gatgatggtc
     3121 tgattaacaa ggaggagttt caacttgctt tgttcaaaag ccggaagagg gagaatatat
     3181 ttgctaatag gatatttgat atgtttgatg ttaaacgaaa aggagtcatc gatttcgggg
     3241 actttgtaag atcactcaat gttttccacc ccaatgcttc tctagaagac aaaatagatt
     3301 ttacgtttag gctttacgac atggactgca cgggttacat tgaacgacaa gaggtcaagc
     3361 aaatgttgat cgcgcttctc tgcgaatctg aaatgaaact ggctgatgaa accatagaga
     3421 taatactcga taagacattt gaagatgcag acgtgaatca agatggaaaa attgataaat
     3481 tagagtggag tgatttcgta aacaaaaatc catcattgct taagatcatg actcttccat
     3541 atctcaggga tattacgaca acgtttccga gttttgtctt ccattcggag gtcgatgaga
     3601 ttgccacata cccagctttc ttgtacaaag tggttgatgg gatggacaag cagaagaacg
     3661 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg
     3721 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3781 acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3841 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat
     3901 ctagagtccg caaaaatcac cagtctctct ctacaaatct atctctctct atttttctcc
     3961 agaataatgt gtgagtagtt cccagataag ggaattaggg ttcttatagg gtttcgctca
     4021 tgtgttgagc atataagaaa cccttagtat gtatttgtat ttgtaaaata cttctatcaa
     4081 taaaatttct aattcctaaa accaaaatcc agtgaccggg cggccgccac cgcggtggag
     4141 ggggatcaga ttgtcgtttc ccgccttcag tttaaactat cagtgtttga caggatatat
     4201 tggcgggtaa acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg
     4261 tgaaaaggtt tatccgttcg tccatttgta tgtgtacatc accgacgagc aaggcaagac
     4321 cgagcgcctt tccgacgctc accgggctgg ttgccctcgc cgctgggctg gcggccgtct
     4381 atggccctgc aaacgcgcca gaaacgccgt cgaagccgtg tgcgagacac cggccgccgg
     4441 cgttgtggat acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac
     4501 acttgagggg ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg
     4561 ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt
     4621 ttcccacaga tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg
     4681 ggcgcgacta ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac
     4741 agatgagggg cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt
     4801 tgcaagggtt tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa
     4861 ccaatattta taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg
     4921 ccgaaggggg gtgccccccc ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc
     4981 cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc
     5041 agtccttgcc attgccggga tcggggcagt aacgggatgg gcgatcagcc cgaaagctac
     5101 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5161 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5221 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5281 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5341 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5401 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5461 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5521 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5581 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5641 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5701 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5761 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5821 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5881 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5941 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6001 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6061 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6121 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6181 ttcgttccac tgagcg
//
