LOCUS       sequence-195327-        6892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  split YFP. Plant expression of CIPK5-YFPc-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6892)
  TITLE     Robatzek Lab plasmids
REFERENCE   2  (bases 1 to 6892)
  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-YFPc-1
FEATURES             Location/Qualifiers
     source          1..6892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     500..519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    911..935
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(1029..1281)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1301..1852)
                     /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(1894..2073)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2542..2886
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2830..2850
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2888..2904
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2889..2905
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2921..2945
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4284..4308)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4319..4570
                     /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     4507..4528
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4862..4886
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4998..5707
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5780..5871
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5872..5940,5941..6732)
                     /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(6090..6109)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
       61 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      121 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg
      661 actctagcta gaggatcaca ggcagcaacg ctctgtcatc gttacaatca acatgctacc
      721 ctccgcgaga tcatccgtgt ttcaaacccg gcagcttagt tgccgttctt ccgaatagca
      781 tcggtaacat gagcaaagtc tgccgcctta caacggctct cccgctgacg ccgtcccgga
      841 ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt
      901 ggctggctgg tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata
      961 acacattgcg gacgttttta atgatcgaat actaacgtct ctaccagata tcagcttgca
     1021 tgccggtcga tctagtaaca tagatgacac cgcgcgcgat aatttatcct agtttgcgcg
     1081 ctatattttg ttttctatcg cgtattaaat gtataattgc gggactctaa tcataaaaac
     1141 ccatctcata aataacgtca tgcattacat gttaattatt acatgcttaa cgtaattcaa
     1201 cagaaattat atgataatca tcgcaagacc ggcaacagga ttcaatctta agaaacttta
     1261 ttgccaaatg tttgaacgat ctgcttgact ctaggggtca tcagatttcg gtgacgggca
     1321 ggaccggacg gggcggcacc ggcaggctga agtccagctg ccagaaaccc acgtcatgcc
     1381 agttcccgtg cttgaagccg gccgcccgca gcatgccgcg gggggcatat ccgagcgcct
     1441 cgtgcatgcg cacgctcggg tcgttgggca gcccgatgac agcgaccacg ctcttgaagc
     1501 cctgtgcctc cagggacttc agcaggtggg tgtagagcgt ggagcccagt cccgtccgct
     1561 ggtggcgggg ggagacgtac acggtcgact cggccgtcca gtcgtaggcg ttgcgtgcct
     1621 tccagggacc cgcgtaggcg atgccggcga cctcgccgtc cacctcggcg acgagccagg
     1681 gatagcgctc ccgcagacgg acgaggtcgt ccgtccactc ctgcggttcc tgcggctcgg
     1741 tacggaagtt gaccgtgctt gtctcgatgt agtggttgac gatggtgcag accgccggca
     1801 tgtccgcctc ggtggcacgg cggatgtcgg ccgggcgtcg ttctgggctc atggtagatc
     1861 ccctcgatcg agttgagagt gaatatgaga ctctaattgg ataccgaggg gaatttatgg
     1921 aacgtcagtg gagcattttt gacaagaaat atttgctagc tgatagtgac cttaggcgac
     1981 ttttgaacgc gcaataatgg tttctgacgt atgtgcttag ctcattaaac tccagaaacc
     2041 cgcggctcag tggctccttc aacgttgcgg ttctgtcagt tccaaacgta aaacggcttg
     2101 tcccgcgtca tcggcggggg tcataacgtg actcccttaa ttctccgctc atgatcagat
     2161 tgtcgtttcc cgccttcggt ttgggcgcgc ccggttcaga agaccagagg gctattgaga
     2221 cttttcaaca aagggtaata tcgggaaacc tcctcggatt ccattgccca gctatctgtc
     2281 acttcatcga aaggacagta gaaaaggaag atggcttcta caaatgccat cattgcgata
     2341 aaggaaaggc tatcgttcaa gatgcctcta ccgacagtgg tcccaaagat ggacccccac
     2401 ccacgaggaa catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt
     2461 gatgtgatac atggtggagc acgacactct cgtctactcc aagaatatca aagatacagt
     2521 ctcagaagac cagagggcta ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct
     2581 cggattccat tgcccagcta tctgtcactt catcgaaagg acagtagaaa aggaagatgg
     2641 cttctacaaa tgccatcatt gcgataaagg aaaggctatc gttcaagatg cctctaccga
     2701 cagtggtccc aaagatggac ccccacccac gaggaacatc gtggaaaaag aagacgttcc
     2761 aaccacgtct tcaaagcaag tggattgatg tgatatctcc actgacgtaa gggatgacgc
     2821 acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga
     2881 gaggacctcg aggtcgacgg tatcgataag cttcccatca acaagtttgt acaaaaaagc
     2941 aggcttgatg gaggaagaac ggcgagttct gttcggaaag tacgagatgg gaaggttatt
     3001 aggcaaagga accttcgcca aagtctatta cggcaaagag atcatcggcg gcgagtgcgt
     3061 cgcaatcaaa gtcatcaaca aggatcaagt aatgaagaga cctggaatga tggaacaaat
     3121 caaacgcgag atttcaatca tgaagctcgt tcgtcatccc aacatagttg aattgaaaga
     3181 agtcatggct acgaagacca agatcttctt cgtcatggag ttcgttaaag gcggcgagct
     3241 tttctgcaaa atctctaaag gtaagttaca cgaagacgct gctcgtagat atttccagca
     3301 attgatctca gccgttgatt attgccatag tagaggcgtt tctcatcgcg atctgaaacc
     3361 tgagaatctt cttttagatg agaatggaga tttgaaaatc tccgatttcg gattatctgc
     3421 gttaccggaa cagattcttc aagacggatt gcttcatacg cagtgtggaa ctccggcgta
     3481 tgttgcgccg gaggtattaa agaagaaagg atacgacggc gcgaaagctg atatctggtc
     3541 gtgcggcgtc gttttgtatg ttcttctcgc cggatgtttg ccgtttcaag atgagaatct
     3601 gatgaatatg tatcggaaga ttttcagagc ggattttgaa tttccgccgt ggttttctcc
     3661 ggaagcgagg aggttgattt cgaaactcct cgttgtagat ccagatcgac ggatctcgat
     3721 tccggcgatt atgagaacgc cttggctccg gaaaaacttc actccgccgt tagctttcaa
     3781 aatcgacgaa ccgatttgtt ctcaaagcag caaaaacaac gaagaagaag aagaagacgg
     3841 agattgtgaa aatcaaacag agccgatatc gccgaaattc ttcaacgctt tcgaattcat
     3901 atcctcgatg tcttccggat ttgacttatc gagcttgttc gagagcaaga ggaaagtgca
     3961 atcggtgttt acgtcacggt cgtcggcgac ggaggtaatg gagaagatag aaacggttac
     4021 gaaggagatg aacatgaaag tgaagagaac gaaggatttt aaagtgaaga tggaagggaa
     4081 aacggaaggg agaaaaggac ggttgtcgat gacggcggaa gtgtttgaag tagcgccgga
     4141 gatatcggtg gtggagtttt gcaaatcggc gggagatact ttggagtacg ataggttata
     4201 cgaagaagaa gtacggccgg cgttaaacga catcgtctgg tcatggcacg gtgataataa
     4261 taacacttct tccgaggatt gttacccagc tttcttgtac aaagtggttg atgggatgga
     4321 caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg
     4381 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     4441 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg
     4501 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     4561 gctgtacaag taatctagag tccgcaaaaa tcaccagtct ctctctacaa atctatctct
     4621 ctctattttt ctccagaata atgtgtgagt agttcccaga taagggaatt agggttctta
     4681 tagggtttcg ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa
     4741 aatacttcta tcaataaaat ttctaattcc taaaaccaaa atccagtgac cgggcggccg
     4801 ccaccgcggt ggagggggat cagattgtcg tttcccgcct tcagtttaaa ctatcagtgt
     4861 ttgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag aataatcgga
     4921 tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgta catcaccgac
     4981 gagcaaggca agaccgagcg cctttccgac gctcaccggg ctggttgccc tcgccgctgg
     5041 gctggcggcc gtctatggcc ctgcaaacgc gccagaaacg ccgtcgaagc cgtgtgcgag
     5101 acaccggccg ccggcgttgt ggatacctcg cggaaaactt ggccctcact gacagatgag
     5161 gggcggacgt tgacacttga ggggccgact cacccggcgc ggcgttgaca gatgaggggc
     5221 aggctcgatt tcggccggcg acgtggagct ggccagcctc gcaaatcggc gaaaacgcct
     5281 gattttacgc gagtttccca cagatgatgt ggacaagcct ggggataagt gccctgcggt
     5341 attgacactt gaggggcgcg actactgaca gatgaggggc gcgatccttg acacttgagg
     5401 ggcagagtgc tgacagatga ggggcgcacc tattgacatt tgaggggctg tccacaggca
     5461 gaaaatccag catttgcaag ggtttccgcc cgtttttcgg ccaccgctaa cctgtctttt
     5521 aacctgcttt taaaccaata tttataaacc ttgtttttaa ccagggctgc gccctgtgcg
     5581 cgtgaccgcg cacgccgaag gggggtgccc ccccttctcg aaccctcccg gcccgctaac
     5641 gcgggcctcc catcccccca ggggctgcgc ccctcggccg cgaacggcct caccccaaaa
     5701 atggcagcgc tggcagtcct tgccattgcc gggatcgggg cagtaacggg atgggcgatc
     5761 agcccgaaag ctacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5821 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5881 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5941 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6001 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6061 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6121 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6181 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6241 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6301 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6361 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6421 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6481 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6541 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6601 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6661 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6721 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6781 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6841 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat ca
//
