LOCUS       sequence-196617-        7102 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  split YFP. Plant expression of CIPK5-YFPn-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7102)
  TITLE     Robatzek Lab plasmids
REFERENCE   2  (bases 1 to 7102)
  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-CIPK5-YFPn-1
FEATURES             Location/Qualifiers
     source          1..7102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    917..941
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(1035..1287)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1307..1858)
                     /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(1900..2079)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2548..2892
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2836..2856
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2894..2910
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2895..2911
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2927..2951
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4290..4314)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(4322..4324,4325..4327,4328..4786)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMA"
     primer_bind     complement(4367..4388)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4628..4647)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    5078..5102
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      5214..5923
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5996..6087
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6088..6156,6157..6948)
                     /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(6306..6325)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      661 acatggactc tagctagagg atcacaggca gcaacgctct gtcatcgtta caatcaacat
      721 gctaccctcc gcgagatcat ccgtgtttca aacccggcag cttagttgcc gttcttccga
      781 atagcatcgg taacatgagc aaagtctgcc gccttacaac ggctctcccg ctgacgccgt
      841 cccggactga tgggctgcct gtatcgagtg gtgattttgt gccgagctgc cggtcgggga
      901 gctgttggct ggctggtggc aggatatatt gtggtgtaaa caaattgacg cttagacaac
      961 ttaataacac attgcggacg tttttaatga tcgaatacta acgtctctac cagatatcag
     1021 cttgcatgcc ggtcgatcta gtaacataga tgacaccgcg cgcgataatt tatcctagtt
     1081 tgcgcgctat attttgtttt ctatcgcgta ttaaatgtat aattgcggga ctctaatcat
     1141 aaaaacccat ctcataaata acgtcatgca ttacatgtta attattacat gcttaacgta
     1201 attcaacaga aattatatga taatcatcgc aagaccggca acaggattca atcttaagaa
     1261 actttattgc caaatgtttg aacgatctgc ttgactctag gggtcatcag atttcggtga
     1321 cgggcaggac cggacggggc ggcaccggca ggctgaagtc cagctgccag aaacccacgt
     1381 catgccagtt cccgtgcttg aagccggccg cccgcagcat gccgcggggg gcatatccga
     1441 gcgcctcgtg catgcgcacg ctcgggtcgt tgggcagccc gatgacagcg accacgctct
     1501 tgaagccctg tgcctccagg gacttcagca ggtgggtgta gagcgtggag cccagtcccg
     1561 tccgctggtg gcggggggag acgtacacgg tcgactcggc cgtccagtcg taggcgttgc
     1621 gtgccttcca gggacccgcg taggcgatgc cggcgacctc gccgtccacc tcggcgacga
     1681 gccagggata gcgctcccgc agacggacga ggtcgtccgt ccactcctgc ggttcctgcg
     1741 gctcggtacg gaagttgacc gtgcttgtct cgatgtagtg gttgacgatg gtgcagaccg
     1801 ccggcatgtc cgcctcggtg gcacggcgga tgtcggccgg gcgtcgttct gggctcatgg
     1861 tagatcccct cgatcgagtt gagagtgaat atgagactct aattggatac cgaggggaat
     1921 ttatggaacg tcagtggagc atttttgaca agaaatattt gctagctgat agtgacctta
     1981 ggcgactttt gaacgcgcaa taatggtttc tgacgtatgt gcttagctca ttaaactcca
     2041 gaaacccgcg gctcagtggc tccttcaacg ttgcggttct gtcagttcca aacgtaaaac
     2101 ggcttgtccc gcgtcatcgg cgggggtcat aacgtgactc ccttaattct ccgctcatga
     2161 tcagattgtc gtttcccgcc ttcggtttgg gcgcgcccgg ttcagaagac cagagggcta
     2221 ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct cggattccat tgcccagcta
     2281 tctgtcactt catcgaaagg acagtagaaa aggaagatgg cttctacaaa tgccatcatt
     2341 gcgataaagg aaaggctatc gttcaagatg cctctaccga cagtggtccc aaagatggac
     2401 ccccacccac gaggaacatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag
     2461 tggattgatg tgatacatgg tggagcacga cactctcgtc tactccaaga atatcaaaga
     2521 tacagtctca gaagaccaga gggctattga gacttttcaa caaagggtaa tatcgggaaa
     2581 cctcctcgga ttccattgcc cagctatctg tcacttcatc gaaaggacag tagaaaagga
     2641 agatggcttc tacaaatgcc atcattgcga taaaggaaag gctatcgttc aagatgcctc
     2701 taccgacagt ggtcccaaag atggaccccc acccacgagg aacatcgtgg aaaaagaaga
     2761 cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat atctccactg acgtaaggga
     2821 tgacgcacaa tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca
     2881 tttggagagg acctcgaggt cgacggtatc gataagcttc ccatcaacaa gtttgtacaa
     2941 aaaagcaggc ttgatggagg aagaacggcg agttctgttc ggaaagtacg agatgggaag
     3001 gttattaggc aaaggaacct tcgccaaagt ctattacggc aaagagatca tcggcggcga
     3061 gtgcgtcgca atcaaagtca tcaacaagga tcaagtaatg aagagacctg gaatgatgga
     3121 acaaatcaaa cgcgagattt caatcatgaa gctcgttcgt catcccaaca tagttgaatt
     3181 gaaagaagtc atggctacga agaccaagat cttcttcgtc atggagttcg ttaaaggcgg
     3241 cgagcttttc tgcaaaatct ctaaaggtaa gttacacgaa gacgctgctc gtagatattt
     3301 ccagcaattg atctcagccg ttgattattg ccatagtaga ggcgtttctc atcgcgatct
     3361 gaaacctgag aatcttcttt tagatgagaa tggagatttg aaaatctccg atttcggatt
     3421 atctgcgtta ccggaacaga ttcttcaaga cggattgctt catacgcagt gtggaactcc
     3481 ggcgtatgtt gcgccggagg tattaaagaa gaaaggatac gacggcgcga aagctgatat
     3541 ctggtcgtgc ggcgtcgttt tgtatgttct tctcgccgga tgtttgccgt ttcaagatga
     3601 gaatctgatg aatatgtatc ggaagatttt cagagcggat tttgaatttc cgccgtggtt
     3661 ttctccggaa gcgaggaggt tgatttcgaa actcctcgtt gtagatccag atcgacggat
     3721 ctcgattccg gcgattatga gaacgccttg gctccggaaa aacttcactc cgccgttagc
     3781 tttcaaaatc gacgaaccga tttgttctca aagcagcaaa aacaacgaag aagaagaaga
     3841 agacggagat tgtgaaaatc aaacagagcc gatatcgccg aaattcttca acgctttcga
     3901 attcatatcc tcgatgtctt ccggatttga cttatcgagc ttgttcgaga gcaagaggaa
     3961 agtgcaatcg gtgtttacgt cacggtcgtc ggcgacggag gtaatggaga agatagaaac
     4021 ggttacgaag gagatgaaca tgaaagtgaa gagaacgaag gattttaaag tgaagatgga
     4081 agggaaaacg gaagggagaa aaggacggtt gtcgatgacg gcggaagtgt ttgaagtagc
     4141 gccggagata tcggtggtgg agttttgcaa atcggcggga gatactttgg agtacgatag
     4201 gttatacgaa gaagaagtac ggccggcgtt aaacgacatc gtctggtcat ggcacggtga
     4261 taataataac acttcttccg aggattgtta cccagctttc ttgtacaaag tggttgatgg
     4321 gatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     4381 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     4441 cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac
     4501 cctcgtgacc accctgggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa
     4561 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     4621 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     4681 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     4741 caagctggag tacaactaca acagccacaa cgtctatatc atggcctaat ctagagtccg
     4801 caaaaatcac cagtctctct ctacaaatct atctctctct atttttctcc agaataatgt
     4861 gtgagtagtt cccagataag ggaattaggg ttcttatagg gtttcgctca tgtgttgagc
     4921 atataagaaa cccttagtat gtatttgtat ttgtaaaata cttctatcaa taaaatttct
     4981 aattcctaaa accaaaatcc agtgaccggg cggccgccac cgcggtggag ggggatcaga
     5041 ttgtcgtttc ccgccttcag tttaaactat cagtgtttga caggatatat tggcgggtaa
     5101 acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt
     5161 tatccgttcg tccatttgta tgtgtacatc accgacgagc aaggcaagac cgagcgcctt
     5221 tccgacgctc accgggctgg ttgccctcgc cgctgggctg gcggccgtct atggccctgc
     5281 aaacgcgcca gaaacgccgt cgaagccgtg tgcgagacac cggccgccgg cgttgtggat
     5341 acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac acttgagggg
     5401 ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg ccggcgacgt
     5461 ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt ttcccacaga
     5521 tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg ggcgcgacta
     5581 ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac agatgagggg
     5641 cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt tgcaagggtt
     5701 tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa ccaatattta
     5761 taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg ccgaaggggg
     5821 gtgccccccc ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc cccccagggg
     5881 ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc agtccttgcc
     5941 attgccggga tcggggcagt aacgggatgg gcgatcagcc cgaaagctac ccctatttgt
     6001 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6061 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6121 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6181 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6241 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6301 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6361 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6421 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6481 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6541 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6601 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6661 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6721 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6781 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6841 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6901 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6961 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7021 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7081 tgagcgtcag accccgtaga aa
//
