LOCUS       sequence-197841-        6223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  split YFP. Plant expression of CBL4-YFPc-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6223)
  TITLE     Robatzek Lab plasmids
REFERENCE   2  (bases 1 to 6223)
  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-CBL4-YFPc-1
FEATURES             Location/Qualifiers
     source          1..6223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      29..617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     518..537
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    929..953
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(1047..1299)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1319..1870)
                     /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(1912..2091)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2560..2904
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2848..2868
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2906..2922
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2907..2923
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2939..2963
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3633..3657)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3668..3919
                     /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     3856..3877
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4211..4235
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4347..5056
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5129..5220
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5221..5289,5290..6081)
                     /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(5439..5458)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
       61 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
      121 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
      181 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
      241 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
      301 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
      361 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
      421 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
      481 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
      541 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
      601 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
      661 ggccttttgc tcacatggac tctagctaga ggatcacagg cagcaacgct ctgtcatcgt
      721 tacaatcaac atgctaccct ccgcgagatc atccgtgttt caaacccggc agcttagttg
      781 ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca acggctctcc
      841 cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt gtgccgagct
      901 gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta aacaaattga
      961 cgcttagaca acttaataac acattgcgga cgtttttaat gatcgaatac taacgtctct
     1021 accagatatc agcttgcatg ccggtcgatc tagtaacata gatgacaccg cgcgcgataa
     1081 tttatcctag tttgcgcgct atattttgtt ttctatcgcg tattaaatgt ataattgcgg
     1141 gactctaatc ataaaaaccc atctcataaa taacgtcatg cattacatgt taattattac
     1201 atgcttaacg taattcaaca gaaattatat gataatcatc gcaagaccgg caacaggatt
     1261 caatcttaag aaactttatt gccaaatgtt tgaacgatct gcttgactct aggggtcatc
     1321 agatttcggt gacgggcagg accggacggg gcggcaccgg caggctgaag tccagctgcc
     1381 agaaacccac gtcatgccag ttcccgtgct tgaagccggc cgcccgcagc atgccgcggg
     1441 gggcatatcc gagcgcctcg tgcatgcgca cgctcgggtc gttgggcagc ccgatgacag
     1501 cgaccacgct cttgaagccc tgtgcctcca gggacttcag caggtgggtg tagagcgtgg
     1561 agcccagtcc cgtccgctgg tggcgggggg agacgtacac ggtcgactcg gccgtccagt
     1621 cgtaggcgtt gcgtgccttc cagggacccg cgtaggcgat gccggcgacc tcgccgtcca
     1681 cctcggcgac gagccaggga tagcgctccc gcagacggac gaggtcgtcc gtccactcct
     1741 gcggttcctg cggctcggta cggaagttga ccgtgcttgt ctcgatgtag tggttgacga
     1801 tggtgcagac cgccggcatg tccgcctcgg tggcacggcg gatgtcggcc gggcgtcgtt
     1861 ctgggctcat ggtagatccc ctcgatcgag ttgagagtga atatgagact ctaattggat
     1921 accgagggga atttatggaa cgtcagtgga gcatttttga caagaaatat ttgctagctg
     1981 atagtgacct taggcgactt ttgaacgcgc aataatggtt tctgacgtat gtgcttagct
     2041 cattaaactc cagaaacccg cggctcagtg gctccttcaa cgttgcggtt ctgtcagttc
     2101 caaacgtaaa acggcttgtc ccgcgtcatc ggcgggggtc ataacgtgac tcccttaatt
     2161 ctccgctcat gatcagattg tcgtttcccg ccttcggttt gggcgcgccc ggttcagaag
     2221 accagagggc tattgagact tttcaacaaa gggtaatatc gggaaacctc ctcggattcc
     2281 attgcccagc tatctgtcac ttcatcgaaa ggacagtaga aaaggaagat ggcttctaca
     2341 aatgccatca ttgcgataaa ggaaaggcta tcgttcaaga tgcctctacc gacagtggtc
     2401 ccaaagatgg acccccaccc acgaggaaca tcgtggaaaa agaagacgtt ccaaccacgt
     2461 cttcaaagca agtggattga tgtgatacat ggtggagcac gacactctcg tctactccaa
     2521 gaatatcaaa gatacagtct cagaagacca gagggctatt gagacttttc aacaaagggt
     2581 aatatcggga aacctcctcg gattccattg cccagctatc tgtcacttca tcgaaaggac
     2641 agtagaaaag gaagatggct tctacaaatg ccatcattgc gataaaggaa aggctatcgt
     2701 tcaagatgcc tctaccgaca gtggtcccaa agatggaccc ccacccacga ggaacatcgt
     2761 ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg atatctccac
     2821 tgacgtaagg gatgacgcac aatcccacta tccttcgcaa gacccttcct ctatataagg
     2881 aagttcattt catttggaga ggacctcgag gtcgacggta tcgataagct tcccatcaac
     2941 aagtttgtac aaaaaagcag gcttgatggg ctgctctgta tcgaagaaga agaagaagaa
     3001 tgcaatgcga ccaccgggat atgaggatcc cgagcttctt gcatccgtca cgccattcac
     3061 ggtagaagaa gtggaggctt tgtatgaact gttcaagaag ctaagcagct caattatcga
     3121 tgatggtctt attcataagg aagaatttca gctggcttta ttcagaaata ggaaccggag
     3181 gaatctcttc gctgatcgga tatttgatgt atttgatgtg aagcgaaatg gagtgatcga
     3241 gtttggtgaa tttgtccggt ccttaggtgt cttccatcca agcgcgccgg tccatgaaaa
     3301 agtcaaattt gctttcaagt tgtacgattt acgacaaact ggattcatcg agcgagaaga
     3361 attgaaagag atggtagtag cgcttcttca cgaatccgaa ctagttcttt ccgaagatat
     3421 gattgaagta atggtggata aggctttcgt gcaagcagac cgcaaaaacg acggaaaaat
     3481 cgatatagat gaatggaaag actttgtatc cttgaatcca tcgctcatca agaacatgac
     3541 tttgccatat ctaaaggaca taaataggac gtttccaagt ttcgtttcat cttgtgaaga
     3601 ggaagaaatg gaattgcaaa acgtatcttc ctacccagct ttcttgtaca aagtggttga
     3661 tgggatggac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
     3721 ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3781 cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa
     3841 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3901 catggacgag ctgtacaagt aatctagagt ccgcaaaaat caccagtctc tctctacaaa
     3961 tctatctctc tctatttttc tccagaataa tgtgtgagta gttcccagat aagggaatta
     4021 gggttcttat agggtttcgc tcatgtgttg agcatataag aaacccttag tatgtatttg
     4081 tatttgtaaa atacttctat caataaaatt tctaattcct aaaaccaaaa tccagtgacc
     4141 gggcggccgc caccgcggtg gagggggatc agattgtcgt ttcccgcctt cagtttaaac
     4201 tatcagtgtt tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga
     4261 ataatcggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgtac
     4321 atcaccgacg agcaaggcaa gaccgagcgc ctttccgacg ctcaccgggc tggttgccct
     4381 cgccgctggg ctggcggccg tctatggccc tgcaaacgcg ccagaaacgc cgtcgaagcc
     4441 gtgtgcgaga caccggccgc cggcgttgtg gatacctcgc ggaaaacttg gccctcactg
     4501 acagatgagg ggcggacgtt gacacttgag gggccgactc acccggcgcg gcgttgacag
     4561 atgaggggca ggctcgattt cggccggcga cgtggagctg gccagcctcg caaatcggcg
     4621 aaaacgcctg attttacgcg agtttcccac agatgatgtg gacaagcctg gggataagtg
     4681 ccctgcggta ttgacacttg aggggcgcga ctactgacag atgaggggcg cgatccttga
     4741 cacttgaggg gcagagtgct gacagatgag gggcgcacct attgacattt gaggggctgt
     4801 ccacaggcag aaaatccagc atttgcaagg gtttccgccc gtttttcggc caccgctaac
     4861 ctgtctttta acctgctttt aaaccaatat ttataaacct tgtttttaac cagggctgcg
     4921 ccctgtgcgc gtgaccgcgc acgccgaagg ggggtgcccc cccttctcga accctcccgg
     4981 cccgctaacg cgggcctccc atccccccag gggctgcgcc cctcggccgc gaacggcctc
     5041 accccaaaaa tggcagcgct ggcagtcctt gccattgccg ggatcggggc agtaacggga
     5101 tgggcgatca gcccgaaagc tacccctatt tgtttatttt tctaaataca ttcaaatatg
     5161 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5221 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5281 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5341 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5401 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5461 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5521 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5581 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5641 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5701 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5761 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5821 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5881 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5941 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6001 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6061 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6121 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6181 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cag
//
