LOCUS       sequence-197551-        6199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  split YFP. Plant expression of CBL8-YFPc-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6199)
  TITLE     Robatzek Lab plasmids
REFERENCE   2  (bases 1 to 6199)
  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-CBL8-YFPc-1
FEATURES             Location/Qualifiers
     source          1..6199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     496..515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    907..931
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(1025..1277)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1297..1848)
                     /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(1890..2069)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2538..2882
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2826..2846
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2884..2900
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2885..2901
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2917..2941
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3587..3611)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3622..3873
                     /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     3810..3831
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4165..4189
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4301..5010
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5083..5174
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5175..5243,5244..6035)
                     /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(5393..5412)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
       61 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      121 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      481 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      541 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      601 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggactc
      661 tagctagagg atcacaggca gcaacgctct gtcatcgtta caatcaacat gctaccctcc
      721 gcgagatcat ccgtgtttca aacccggcag cttagttgcc gttcttccga atagcatcgg
      781 taacatgagc aaagtctgcc gccttacaac ggctctcccg ctgacgccgt cccggactga
      841 tgggctgcct gtatcgagtg gtgattttgt gccgagctgc cggtcgggga gctgttggct
      901 ggctggtggc aggatatatt gtggtgtaaa caaattgacg cttagacaac ttaataacac
      961 attgcggacg tttttaatga tcgaatacta acgtctctac cagatatcag cttgcatgcc
     1021 ggtcgatcta gtaacataga tgacaccgcg cgcgataatt tatcctagtt tgcgcgctat
     1081 attttgtttt ctatcgcgta ttaaatgtat aattgcggga ctctaatcat aaaaacccat
     1141 ctcataaata acgtcatgca ttacatgtta attattacat gcttaacgta attcaacaga
     1201 aattatatga taatcatcgc aagaccggca acaggattca atcttaagaa actttattgc
     1261 caaatgtttg aacgatctgc ttgactctag gggtcatcag atttcggtga cgggcaggac
     1321 cggacggggc ggcaccggca ggctgaagtc cagctgccag aaacccacgt catgccagtt
     1381 cccgtgcttg aagccggccg cccgcagcat gccgcggggg gcatatccga gcgcctcgtg
     1441 catgcgcacg ctcgggtcgt tgggcagccc gatgacagcg accacgctct tgaagccctg
     1501 tgcctccagg gacttcagca ggtgggtgta gagcgtggag cccagtcccg tccgctggtg
     1561 gcggggggag acgtacacgg tcgactcggc cgtccagtcg taggcgttgc gtgccttcca
     1621 gggacccgcg taggcgatgc cggcgacctc gccgtccacc tcggcgacga gccagggata
     1681 gcgctcccgc agacggacga ggtcgtccgt ccactcctgc ggttcctgcg gctcggtacg
     1741 gaagttgacc gtgcttgtct cgatgtagtg gttgacgatg gtgcagaccg ccggcatgtc
     1801 cgcctcggtg gcacggcgga tgtcggccgg gcgtcgttct gggctcatgg tagatcccct
     1861 cgatcgagtt gagagtgaat atgagactct aattggatac cgaggggaat ttatggaacg
     1921 tcagtggagc atttttgaca agaaatattt gctagctgat agtgacctta ggcgactttt
     1981 gaacgcgcaa taatggtttc tgacgtatgt gcttagctca ttaaactcca gaaacccgcg
     2041 gctcagtggc tccttcaacg ttgcggttct gtcagttcca aacgtaaaac ggcttgtccc
     2101 gcgtcatcgg cgggggtcat aacgtgactc ccttaattct ccgctcatga tcagattgtc
     2161 gtttcccgcc ttcggtttgg gcgcgcccgg ttcagaagac cagagggcta ttgagacttt
     2221 tcaacaaagg gtaatatcgg gaaacctcct cggattccat tgcccagcta tctgtcactt
     2281 catcgaaagg acagtagaaa aggaagatgg cttctacaaa tgccatcatt gcgataaagg
     2341 aaaggctatc gttcaagatg cctctaccga cagtggtccc aaagatggac ccccacccac
     2401 gaggaacatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg
     2461 tgatacatgg tggagcacga cactctcgtc tactccaaga atatcaaaga tacagtctca
     2521 gaagaccaga gggctattga gacttttcaa caaagggtaa tatcgggaaa cctcctcgga
     2581 ttccattgcc cagctatctg tcacttcatc gaaaggacag tagaaaagga agatggcttc
     2641 tacaaatgcc atcattgcga taaaggaaag gctatcgttc aagatgcctc taccgacagt
     2701 ggtcccaaag atggaccccc acccacgagg aacatcgtgg aaaaagaaga cgttccaacc
     2761 acgtcttcaa agcaagtgga ttgatgtgat atctccactg acgtaaggga tgacgcacaa
     2821 tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagagg
     2881 acctcgaggt cgacggtatc gataagcttc ccatcaacaa gtttgtacaa aaaagcaggc
     2941 ttgatgttgg cattcgtgaa atgcttctcg ttgaagcgag caaagcatcc tcgtggatat
     3001 gaggatcctc atgttcttgc atccgaaact cccttcaccg tgaatgagat tgaggcttta
     3061 cacgatttat tcaagaagct aagtacatct attatcaatg atggtctaat ccataaggaa
     3121 gagtttcttc tggctttgtt cagaaacggc agtatgcaaa atctatttgc ggacagggtg
     3181 ttttatatgt ttgataggaa acgcaacggt gtgattgagt ttggagaatt tgttcgttcc
     3241 cttagcatat tccatccata cactcccgaa catgaaaaat ctgcatttat gttcaagctt
     3301 tttgacctcc atggaaccgg cttcatcgag ccccatgagc tgaagaagat ggttggcgca
     3361 ctacttggtg aaacagactt agaactctcg gaagaatcta ttgaagctat cgtggaacag
     3421 acgatgctcg aggttgatac aaacaaagat gggaagattg atgaagaaga gtggaaagag
     3481 ctcgttgcta agaatccttc aatcctcaag aacatgactc tgccttacct caaggaagtg
     3541 actttagcat ttccaagctt tgtactcgac tctgaagttg aagactaccc agctttcttg
     3601 tacaaagtgg ttgatgggat ggacaagcag aagaacggca tcaaggccaa cttcaagatc
     3661 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     3721 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg
     3781 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3841 gggatcactc tcggcatgga cgagctgtac aagtaatcta gagtccgcaa aaatcaccag
     3901 tctctctcta caaatctatc tctctctatt tttctccaga ataatgtgtg agtagttccc
     3961 agataaggga attagggttc ttatagggtt tcgctcatgt gttgagcata taagaaaccc
     4021 ttagtatgta tttgtatttg taaaatactt ctatcaataa aatttctaat tcctaaaacc
     4081 aaaatccagt gaccgggcgg ccgccaccgc ggtggagggg gatcagattg tcgtttcccg
     4141 ccttcagttt aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa
     4201 gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc
     4261 atttgtatgt gtacatcacc gacgagcaag gcaagaccga gcgcctttcc gacgctcacc
     4321 gggctggttg ccctcgccgc tgggctggcg gccgtctatg gccctgcaaa cgcgccagaa
     4381 acgccgtcga agccgtgtgc gagacaccgg ccgccggcgt tgtggatacc tcgcggaaaa
     4441 cttggccctc actgacagat gaggggcgga cgttgacact tgaggggccg actcacccgg
     4501 cgcggcgttg acagatgagg ggcaggctcg atttcggccg gcgacgtgga gctggccagc
     4561 ctcgcaaatc ggcgaaaacg cctgatttta cgcgagtttc ccacagatga tgtggacaag
     4621 cctggggata agtgccctgc ggtattgaca cttgaggggc gcgactactg acagatgagg
     4681 ggcgcgatcc ttgacacttg aggggcagag tgctgacaga tgaggggcgc acctattgac
     4741 atttgagggg ctgtccacag gcagaaaatc cagcatttgc aagggtttcc gcccgttttt
     4801 cggccaccgc taacctgtct tttaacctgc ttttaaacca atatttataa accttgtttt
     4861 taaccagggc tgcgccctgt gcgcgtgacc gcgcacgccg aaggggggtg cccccccttc
     4921 tcgaaccctc ccggcccgct aacgcgggcc tcccatcccc ccaggggctg cgcccctcgg
     4981 ccgcgaacgg cctcacccca aaaatggcag cgctggcagt ccttgccatt gccgggatcg
     5041 gggcagtaac gggatgggcg atcagcccga aagctacccc tatttgttta tttttctaaa
     5101 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5161 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5221 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5281 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5341 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5401 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5461 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5521 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5581 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5641 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5701 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5761 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5821 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5881 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5941 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6001 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6061 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6121 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6181 ccgtagaaaa gatcaaagg
//
