LOCUS       sequence-361300-        4081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of KO1 in plant cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4081)
  AUTHORS   Fujii Y, Yoshimura A, Kodama Y
  TITLE     A novel orange-colored bimolecular fluorescence complementation 
            (BiFC) assay using monomeric Kusabira-Orange protein.
  JOURNAL   Biotechniques. 2018 Apr;64(4):153-161. doi: 10.2144/btn-2017-0121.
  PUBMED    29661017
REFERENCE   2  (bases 1 to 4081)
  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"; doi: "10"; journalName: 
            "Biotechniques"; date: "2018-04"; volume: "64"; issue: "4"; pages: 
            "153-161"
COMMENT     Sequence Label: p35S-KO1
FEATURES             Location/Qualifiers
     source          1..4081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     60..78
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(116..138)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     238..257
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     451..473
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     465..482
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     466..482
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        542..887
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     830..850
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    907..962
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             982..1638
                     /codon_start=1
                     /product="Kusabira-Orange fluorescent protein (Karasawa et 
                     al., 2004)"
                     /label=Kusabira-Orange
                     /translation="MVSVIKPEMKMKYFMDGSVNGHEFTVEGEGTGKPYEGHQEMTLRV
                     TMAKGGPMPFSFDLVSHTFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLQFEDGGFA
                     AVSAHISLRGNCFEHKSKFVGVNFPADGPVMQNQSSDWEPSTEKITTCDGVLKGDVTMY
                     LKLAGGGNHKCQFKTTYKAAKKILKMPQSHFIGHRLVRKTEGNITELVEDAVAHC"
     terminator      1657..1909
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1947..1963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1947..1963)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1960..1982)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1971..1987
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1995..2001,2002..2019,2020..2025))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2040..2061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2178..2195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2349..2937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2429..2448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3108..3899,3900..3968))
                     /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     3731..3750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3969..4073)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
       61 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
      121 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
      181 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
      241 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
      301 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
      361 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      421 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      481 gtgccaagct tgcatgcctg caggctctag aggatccccc ctcagaagac cagagggcta
      541 ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct cggattccat tgcccagcta
      601 tctgtcactt catcgaaagg acagtagaaa aggaaggtgg ctcctacaaa tgccatcatt
      661 gcgataaagg aaaggctatc gttcaagatg cctctaccga cagtggtccc aaagatggac
      721 ccccacccac gaggaacatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag
      781 tggattgatg tgatatctcc actgacgtaa gggatgacgc acaatcccac tatccttcgc
      841 aagacccttc ctctatataa ggaagttcat ttcatttgga gaggacaggc ttcttgagat
      901 ccttcaacaa ttaccaacaa caacaaacaa caaacaacat tacaattact atttacaatt
      961 acagtcgact ctagaggatc catggtgagt gtgattaaac cagagatgaa gatgaagtac
     1021 ttcatggacg gctccgtcaa tgggcatgag ttcacagttg aaggtgaagg cacaggcaaa
     1081 ccttacgagg gacatcaaga gatgacacta cgcgtcacaa tggccaaggg cgggccaatg
     1141 cctttctcgt ttgacttagt gtcacacacg ttctgttacg gccacagacc ttttactaaa
     1201 tatccagaag agataccaga ctatttcaaa caagcatttc ctgaaggcct gtcatgggaa
     1261 aggtcgttgc agttcgaaga tggtgggttc gctgccgtca gtgcgcatat aagccttaga
     1321 ggaaactgct tcgagcacaa atccaaattt gttggggtta actttcctgc cgatggtcct
     1381 gtgatgcaaa accaaagttc tgattgggag ccatcaaccg agaaaattac tacctgcgac
     1441 ggagttctga agggtgatgt tacgatgtac ctaaaacttg cgggaggcgg caatcacaaa
     1501 tgccaattca agactactta caaggcggca aaaaagattc ttaaaatgcc acaaagccat
     1561 ttcatcgggc atcgcctcgt caggaaaacc gaaggcaaca ttactgagct ggtagaagat
     1621 gcagtagctc attgctaaag cggccgcccg gctgcagatc gttcaaacat ttggcaataa
     1681 agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg
     1741 aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt
     1801 tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc
     1861 gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcc gatgataagc
     1921 tgtcaaacat gagaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg
     1981 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2041 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2101 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2161 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2221 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2281 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2341 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2401 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2461 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2521 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2581 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2641 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2701 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2761 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2821 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2881 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2941 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3001 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3061 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3121 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3181 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3241 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3301 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3361 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3421 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3481 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3541 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3601 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3661 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3721 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3781 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3841 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3901 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3961 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4021 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4081 c
//
