LOCUS       62056_17765        9874 bp DNA     circular SYN 04-SEP-2019
DEFINITION  Plant binary expression vector pNC-BiFC-ED2, complete sequence.
ACCESSION   MK896891
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9874)
  AUTHORS   Yan P, Zeng Y, Shen W, Tuo D, Li X, Zhou P.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-MAY-2019) Institute of Tropical Bioscience and 
            Biotechnology, Chinese Academy of Tropical Agriculture Sciences, 
            Xueyuan Road, Haikou, Hainan 571101, China
REFERENCE   2  (bases 1 to 9874)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (07-MAY-2019) Institute of Tropical Bioscience and Biotechnology, 
            Chinese Academy of Tropical Agriculture Sciences, Xueyuan Road, 
            Haikou, Hainan 571101, China"
FEATURES             Location/Qualifiers
     source          1..9874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..26
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        118..789
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     regulatory      799..929
                     /label=translational enhancer
                     /note="translational enhancer"
                     /regulatory_class="enhancer"
     CDS             932..1450
                     /label=VN173
                     /note="N-terminal fragment of mVenus for use in bimolecular
                     fluorescence complementation (BiFC) (Kodama and Hu, 2010)"
     promoter        1650..1752
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             1783..2085
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     regulatory      2155..2365
                     /label=CaMV 35S terminator
                     /note="CaMV 35S terminator"
                     /regulatory_class="terminator"
     CDS             complement(2475..3128)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     regulatory      3227..3437
                     /label=CaMV 35S terminator
                     /note="CaMV 35S terminator"
                     /regulatory_class="terminator"
     gene            3459..3653
                     /gene="CEYFP"
                     /label=CEYFP
     CDS             complement(3714..4016)
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     promoter        complement(4047..4149)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     regulatory      4332..4462
                     /label=translational enhancer
                     /note="translational enhancer"
                     /regulatory_class="enhancer"
     promoter        complement(4472..5143)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     primer_bind     complement(5235..5251)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5274..5290)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5327..5345)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5366..5382)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5390..5406)
                     /label=lac operator
                     /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(5414..5445)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5460..5481)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    5720..5744
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5835..6423)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6527..7339)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      7630..8065
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    8198..8220
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     terminator      complement(8244..8495)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(8538..9329)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(9363..9542)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        9812..9830
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     9856..9872
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagcttgttg taaaacgacg gccagtgccg gcgcgccacc ggtcaacatg tggagcacga
       61 cacacttgtc tactccaaaa atatcaaaga tacagtctca gaagaccaaa gggcaattga
      121 gacttttcaa caaagggtaa tatccggaaa cctcctcgga ttccattgcc cagctatctg
      181 tcactttatt gtgaagatag tggaaaagga aggtggctcc tacaaatgcc atcattgcga
      241 taaaggaaag gccatcgttg aagatgcctc tgccgacagt ggtcccaaag atggaccccc
      301 acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga
      361 ttgatgtgat aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac
      421 agtctcagaa gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct
      481 cctcggattc cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg
      541 tggctcctac aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc
      601 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt
      661 tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga
      721 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
      781 ggagaggacg tcgagagttc tcaacacaac atatacaaaa caaacgaatc tcaagcaatc
      841 aagcattcta cttctattgc agcaatttaa atcatttctt ttaaagcaaa agcaattttc
      901 tgaaaatttt caccatttac gaacgatagc catggtgagc aagggcgagg agctgttcac
      961 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1021 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     1081 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca
     1141 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1201 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1261 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1321 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1381 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     1441 caacatcgag gactccggac tcagatctca gtggtctctg tccagtcctg gcctcgtcgg
     1501 ccattctcga ctaagttggc agcatcaccc gacgcacttt gcgccgaata aatacctgtg
     1561 acggaagatc acttcgcaga ataaataaat cctggtgtcc ctgttgatac cgggaagccc
     1621 tgggccaact tttggcgaaa atgagacgtt gatcggcacg taagaggttc caactttcac
     1681 cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct
     1741 aaggaagcta aacttttgct gacgagaaca gggactggtg aaatgcagtt taaggtttac
     1801 acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac
     1861 acgcctgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc
     1921 tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc
     1981 gatatggcca gtgtgccggt atccgttatc ggggaagaag tggctgatct cagccaccgc
     2041 gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc
     2101 cttatacaca gggccagtct ggccacttgt ggtctgctga gaccctgatc tagagtccgc
     2161 aaaaatcacc agtctctctc tacaaatcta tctctctcta tttttctcca gaataatgtg
     2221 tgagtagttc ccagataagg gaattagggt tcttataggg tttcgctcat gtgttgagca
     2281 tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat aaaatttcta
     2341 attcctaaaa ccaaaatcca gtgacgctta tgggaccgac atatcagtat atattcttat
     2401 accgcaaaaa tcagcgcgca aatacgcata ctgttatctg gcttttagta agccggatcc
     2461 acgcggcgtt tacgcccccc ctgccactca tcgcagtact gttgtaattc attaagcatt
     2521 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
     2581 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
     2641 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
     2701 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
     2761 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     2821 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     2881 accagctcac cgtctttcat tgccatacgg aactccggat gagcattcat caggcgggca
     2941 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
     3001 gccgtaatat ccagctgaac ggtctggtta taggtacatt gaacaactga ctgaaatgcc
     3061 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
     3121 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
     3181 agtgatctta tttcattatg gtgaaagttg gaacctctta ccgggcgtca ctggattttg
     3241 gttttaggaa ttagaaattt tattgataga agtattttac aaatacaaat acatactaag
     3301 ggtttcttat atgctcaaca catgagcgaa accctataag aaccctaatt cccttatctg
     3361 ggaactactc acacattatt ctggagaaaa atagagagag atagatttgt agagagagac
     3421 tggtgatttt tgcggactct agatcaggtg gatcttcact tgtacagctc gtccatgccg
     3481 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     3541 gggtctttgc tcagggcgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg
     3601 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccccactgg
     3661 aaacatccag gtctggccag actggccctg tgtataaggg agcctgacat ttatattccc
     3721 cagaacatca ggttaatggc gtttttgatg tcattttcgc ggtggctgag atcagccact
     3781 tcttccccga taacggatac cggcacactg gccatatcgg tggtcatcat gcgccagctt
     3841 tcatccccga tatgcaccac cgggtaaagt tcacgggaga ctttatctga cagcagacgt
     3901 gcactggcca gggggatcac catccgtcgc ccaggcgtgt caataatatc actctgtaca
     3961 tccacaaaca gacgataacg gctctctctt ttataggtgt aaaccttaaa ctgcatttca
     4021 ccagtccctg ttctcgtcag caaaagttta gcttccttag ctcctgaaaa tctcgataac
     4081 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     4141 tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg gcttcccggt atcaacaggg
     4201 acaccaggat ttatttattc tgcgaagtga tcttccgtca caggtattta ttcggcgcaa
     4261 agtgcgtcgg gtgatgctgc caacttagtc gagaatggcc gacgaggcca cgcatggtcg
     4321 agccaactct gctatcgttc gtaaatggtg aaaattttca gaaaattgct tttgctttaa
     4381 aagaaatgat ttaaattgct gcaatagaag tagaatgctt gattgcttga gattcgtttg
     4441 ttttgtatat gttgtgttga gaactctcga cgtcctctcc aaatgaaatg aacttcctta
     4501 tatagaggaa gggtcttgcg aaggatagtg ggattgtgcg tcatccctta cgtcagtgga
     4561 gatatcacat caatccactt gctttgaaga cgtggttgga acgtcttctt tttccacgat
     4621 gctcctcgtg ggtgggggtc catctttggg accactgtcg gcagaggcat cttcaacgat
     4681 ggcctttcct ttatcgcaat gatggcattt gtaggagcca ccttcctttt ccactatctt
     4741 cacaataaag tgacagatag ctgggcaatg gaatccgagg aggtttccgg atattaccct
     4801 ttgttgaaaa gtctcaattg ccctttggtc ttctgagact gtatctttga tatttttgga
     4861 gtagacaagt gtgtcgtgct ccaccatgtt atcacatcaa tccacttgct ttgaagacgt
     4921 ggttggaacg tcttcttttt ccacgatgct cctcgtgggt gggggtccat ctttgggacc
     4981 actgtcggca gaggcatctt caacgatggc ctttccttta tcgcaatgat ggcatttgta
     5041 ggagccacct tccttttcca ctatcttcac aataaagtga cagatagctg ggcaatggaa
     5101 tccgaggagg tttccggata ttaccctttg ttgaaaagtc tcaattgccc tttggtcttc
     5161 tgagactgta tctttgatat ttttggagta gacaagtgtg tcgtgctcca catgttgacc
     5221 ggtggcgcgc cggcactggc cgtcgtttta caacgaattc ctgcagcccg ggggatccac
     5281 tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt
     5341 taattccgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5401 tcacaattcc acacaacata cgagccggaa ghcataaagt gtaaagcctg gggtgcctaa
     5461 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5521 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5581 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5641 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5701 ggaaagaaca tgaaggcctt gacaggatat attggcgggt aaactaagtc gctgtatgtg
     5761 tttgtttgag atctcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5821 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5881 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5941 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6001 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6061 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6121 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6181 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6241 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6301 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6361 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6421 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6481 taagggattt tggtcatggt tacaaccaat taaccaattc tgattagaaa aactcatcga
     6541 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     6601 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     6661 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     6721 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     6781 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     6841 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     6901 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     6961 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     7021 atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     7081 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     7141 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     7201 gcttcccata caatccatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     7261 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctggag caagacgttt
     7321 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     7381 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa
     7441 cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc acgcatcttc
     7501 ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac tggtccacct
     7561 acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat ggggcgattc
     7621 aggcgatccc catccaacag cccgccgtcg agcgggcttt tttatccccg gaagcctgtg
     7681 gatagagggt agttatccac gtgaaaccgc taatgccccg caaagccttg attcacgggg
     7741 ctttccggcc cgctccaaaa actatccacg tgaaatcgct aatcagggta cgtgaaatcg
     7801 ctaatcggag tacgtgaaat cgctaataag gtcacgtgaa atcgctaatc aaaaaggcac
     7861 gtgagaacgc taatagccct ttcagatcaa cagcttgcaa acacccctcg ctccggcaag
     7921 tagttacagc aagtagtatg ttcaattagc ttttcaatta tgaatatata tatcaattat
     7981 tggtcgccct tggcttgtgg acaatgcgct acgcgcaccg gctccgcccg tggacaaccg
     8041 caagcggttg cccaccgtcg agcgccagcg cctttgccca caacccggcg gccggccgca
     8101 acagatcgtt ttataaattt ttttttttga aaaagaaaaa gcccgaaagg cggcaacctc
     8161 tcgggcttct ggatttccga tccccggaat tagatcttgg caggatatat tgtggtgtaa
     8221 cgttatcagc ttgcatgccg gtcgatctag taacatagat gacaccgcgc gcgataattt
     8281 atcctagttt gcgcgctata ttttgttttc tatcgcgtat taaatgtata attgcgggac
     8341 tctaatcaaa aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc
     8401 ttaacgtaat tcaacagaaa ttatatgata atcatcgcaa gaccggcaac aggattcaat
     8461 cttaagaaac tttattgcca aatgtttgaa cgatctgctt gactctagct agagtccgaa
     8521 ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa
     8581 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct
     8641 tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg
     8701 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca
     8761 tcgccctggg tcacgacgag atcctcgccg tcgggcatcc gcgccttgag cctggcgaac
     8821 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg
     8881 gcttccatcc gagtacgtcc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag
     8941 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg
     9001 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag
     9061 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc
     9121 agccacgata gccgcgctgc ctcgtcttgg agttcattca gggcaccgga caggtcggtc
     9181 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag
     9241 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa
     9301 cctgcgtgca atccatcttg ttcaatcatg cctcgatcga gttgagagtg aatatgagac
     9361 tctaattgga taccgagggg aatttatgga acgtcagtgg agcatttttg acaagaaata
     9421 tttgctagct gatagtgacc ttaggcgact tttgaacgcg caataatggt ttctgacgta
     9481 tgtgcttagc tcattaaact ccagaaaccc gcggctgagt ggctccttca acgttgcggt
     9541 tctgtcagtt ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt cataacgtga
     9601 ctcccttaat tctcatgtat cgataacatt aacgtttaca atttcgcgcc attcgccatt
     9661 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     9721 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     9781 acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gcgaattggg
     9841 taccgggccc cccctcgagg tcgacggtat cgat
//
