LOCUS       sequence-196626-        6235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  split YFP. Plant expression of CBL2-YFPc-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6235)
  TITLE     Robatzek Lab plasmids
REFERENCE   2  (bases 1 to 6235)
  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-CBL2-YFPc-1
FEATURES             Location/Qualifiers
     source          1..6235
                     /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(3645..3669)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3680..3931
                     /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     3868..3889
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4223..4247
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4359..5068
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5141..5232
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5233..5301,5302..6093)
                     /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(5451..5470)
                     /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 gcttgatgtc gcagtgcgtt gacggtatca agcatttatg
     3001 cacctctgtt ttagggtgtt ttgatcttga tttgtataaa caatctggtg gccttggaga
     3061 tcctgaattg cttgcaagag acactgtttt tagtgtgagt gaaatagaag ctctttatga
     3121 gctttttaag aaaatcagca gtgccgtgat tgatgatggg ctaataaaca aggaagagtt
     3181 tcagttggct ttgtttaaga caaataaaaa agagagtttg tttgcagatc gggttttcga
     3241 tttgtttgat acaaagcaca atggaatact ggggtttgag gaatttgctc gtgctctctc
     3301 cgtctttcac ccaaatgcgc ctatagatga caaaattcac ttttcatttc agctatatga
     3361 tcttaaacag caagggttta tcgaacggca agaggtgaaa caaatggttg tcgctacgct
     3421 tgctgaatct ggaatgaatt tgaaggatac tgtcattgag gatataatcg acaagacgtt
     3481 tgaggaagca gacacaaaac acgatggaaa gatcgataag gaagagtgga gaagccttgt
     3541 tcttagacat cccagccttc tgaagaacat gactcttcaa tatctcaagg atatcacgac
     3601 tacgtttcca agctttgtgt tccattctca ggttgaagat acctacccag ctttcttgta
     3661 caaagtggtt gatgggatgg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     3721 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
     3781 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag
     3841 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     3901 gatcactctc ggcatggacg agctgtacaa gtaatctaga gtccgcaaaa atcaccagtc
     3961 tctctctaca aatctatctc tctctatttt tctccagaat aatgtgtgag tagttcccag
     4021 ataagggaat tagggttctt atagggtttc gctcatgtgt tgagcatata agaaaccctt
     4081 agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa
     4141 aatccagtga ccgggcggcc gccaccgcgg tggaggggga tcagattgtc gtttcccgcc
     4201 ttcagtttaa actatcagtg tttgacagga tatattggcg ggtaaaccta agagaaaaga
     4261 gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat
     4321 ttgtatgtgt acatcaccga cgagcaaggc aagaccgagc gcctttccga cgctcaccgg
     4381 gctggttgcc ctcgccgctg ggctggcggc cgtctatggc cctgcaaacg cgccagaaac
     4441 gccgtcgaag ccgtgtgcga gacaccggcc gccggcgttg tggatacctc gcggaaaact
     4501 tggccctcac tgacagatga ggggcggacg ttgacacttg aggggccgac tcacccggcg
     4561 cggcgttgac agatgagggg caggctcgat ttcggccggc gacgtggagc tggccagcct
     4621 cgcaaatcgg cgaaaacgcc tgattttacg cgagtttccc acagatgatg tggacaagcc
     4681 tggggataag tgccctgcgg tattgacact tgaggggcgc gactactgac agatgagggg
     4741 cgcgatcctt gacacttgag gggcagagtg ctgacagatg aggggcgcac ctattgacat
     4801 ttgaggggct gtccacaggc agaaaatcca gcatttgcaa gggtttccgc ccgtttttcg
     4861 gccaccgcta acctgtcttt taacctgctt ttaaaccaat atttataaac cttgttttta
     4921 accagggctg cgccctgtgc gcgtgaccgc gcacgccgaa ggggggtgcc cccccttctc
     4981 gaaccctccc ggcccgctaa cgcgggcctc ccatcccccc aggggctgcg cccctcggcc
     5041 gcgaacggcc tcaccccaaa aatggcagcg ctggcagtcc ttgccattgc cgggatcggg
     5101 gcagtaacgg gatgggcgat cagcccgaaa gctaccccta tttgtttatt tttctaaata
     5161 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5221 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5281 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5341 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5401 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5461 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5521 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5581 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5641 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5701 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5761 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5821 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5881 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5941 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6001 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6061 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6121 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6181 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcag
//
