LOCUS       62056_17775        9870 bp DNA     circular SYN 04-SEP-2019
DEFINITION  Plant binary expression vector pNC-BiFC-ED4, complete sequence.
ACCESSION   MK896893
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9870)
  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 9870)
  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..9870
                     /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"
     promoter        1112..1214
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             1245..1547
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             1608..2126
                     /label=VN173
                     /note="N-terminal fragment of mVenus for use in bimolecular
                     fluorescence complementation (BiFC) (Kodama and Hu, 2010)"
     regulatory      2151..2361
                     /label=CaMV 35S terminator
                     /note="CaMV 35S terminator"
                     /regulatory_class="terminator"
     CDS             2460..3113
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     regulatory      3223..3433
                     /label=CaMV 35S terminator
                     /note="CaMV 35S terminator"
                     /regulatory_class="terminator"
     gene            3455..3649
                     /gene="CEYFP"
                     /label=CEYFP
     CDS             complement(3710..4012)
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     promoter        complement(4043..4145)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     regulatory      4328..4458
                     /label=translational enhancer
                     /note="translational enhancer"
                     /regulatory_class="enhancer"
     promoter        complement(4468..5139)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     primer_bind     complement(5231..5247)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5270..5286)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5323..5341)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5362..5378)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5386..5402)
                     /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(5410..5441)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5456..5477)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    5716..5740
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5831..6419)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6523..7335)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      7626..8061
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    8194..8216
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     terminator      complement(8240..8491)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(8534..9325)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(9359..9538)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        9808..9826
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     9852..9868
                     /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 cagtggtctc tgtccagtcc tggcctcgtc
      961 ggccattctc gactaagttg gcagcatcac ccgacgcact ttgcgccgaa taaatacctg
     1021 tgacggaaga tcacttcgca gaataaataa atcctggtgt ccctgttgat accgggaagc
     1081 cctgggccaa cttttggcga aaatgagacg ttgatcggca cgtaagaggt tccaactttc
     1141 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag
     1201 ctaaggaagc taaacttttg ctgacgagaa cagggactgg tgaaatgcag tttaaggttt
     1261 acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg
     1321 acacgcctgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag
     1381 tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca
     1441 ccgatatggc cagtgtgccg gtatccgtta tcggggaaga agtggctgat ctcagccacc
     1501 gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct
     1561 cccttataca cagggccagt ctggccactt gtggtctgct gagacccatg gtgagcaagg
     1621 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1681 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1741 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
     1801 tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1861 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1921 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1981 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2041 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     2101 acttcaagat ccgccacaac atcgaggact gaagatccac ctgatctaga gtccgcaaaa
     2161 atcaccagtc tctctctaca aatctatctc tctctatttt tctccagaat aatgtgtgag
     2221 tagttcccag ataagggaat tagggttctt atagggtttc gctcatgtgt tgagcatata
     2281 agaaaccctt agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc
     2341 ctaaaaccaa aatccagtga cgcccggtaa gaggttccaa ctttcaccat aatgaaataa
     2401 gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa
     2461 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
     2521 attttgaggc atttcagtca gttgttcaat gtacctataa ccagaccgtt cagctggata
     2581 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     2641 acattcttgc ccgcctgatg aatgctcatc cggagttccg tatggcaatg aaagacggtg
     2701 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     2761 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     2821 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     2881 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     2941 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
     3001 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
     3061 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggg gggcgtaaac
     3121 gccgcgtgga tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt
     3181 ttgcggtata agaatatata ctgatatgtc ggtcccataa gcgtcactgg attttggttt
     3241 taggaattag aaattttatt gatagaagta ttttacaaat acaaatacat actaagggtt
     3301 tcttatatgc tcaacacatg agcgaaaccc tataagaacc ctaattccct tatctgggaa
     3361 ctactcacac attattctgg agaaaaatag agagagatag atttgtagag agagactggt
     3421 gatttttgcg gactctagat caggtggatc ttcacttgta cagctcgtcc atgccgagag
     3481 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     3541 ctttgctcag ggcggactgg tagctcaggt agtggttgtc gggcagcagc acggggccgt
     3601 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccc cactggaaac
     3661 atccaggtct ggccagactg gccctgtgta taagggagcc tgacatttat attccccaga
     3721 acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca gccacttctt
     3781 ccccgataac ggataccggc acactggcca tatcggtggt catcatgcgc cagctttcat
     3841 ccccgatatg caccaccggg taaagttcac gggagacttt atctgacagc agacgtgcac
     3901 tggccagggg gatcaccatc cgtcgcccag gcgtgtcaat aatatcactc tgtacatcca
     3961 caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc atttcaccag
     4021 tccctgttct cgtcagcaaa agtttagctt ccttagctcc tgaaaatctc gataactcaa
     4081 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     4141 gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac
     4201 caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg
     4261 cgtcgggtga tgctgccaac ttagtcgaga atggccgacg aggccacgca tggtcgagcc
     4321 aactctgcta tcgttcgtaa atggtgaaaa ttttcagaaa attgcttttg ctttaaaaga
     4381 aatgatttaa attgctgcaa tagaagtaga atgcttgatt gcttgagatt cgtttgtttt
     4441 gtatatgttg tgttgagaac tctcgacgtc ctctccaaat gaaatgaact tccttatata
     4501 gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc agtggagata
     4561 tcacatcaat ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc
     4621 ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag aggcatcttc aacgatggcc
     4681 tttcctttat cgcaatgatg gcatttgtag gagccacctt ccttttccac tatcttcaca
     4741 ataaagtgac agatagctgg gcaatggaat ccgaggaggt ttccggatat taccctttgt
     4801 tgaaaagtct caattgccct ttggtcttct gagactgtat ctttgatatt tttggagtag
     4861 acaagtgtgt cgtgctccac catgttatca catcaatcca cttgctttga agacgtggtt
     4921 ggaacgtctt ctttttccac gatgctcctc gtgggtgggg gtccatcttt gggaccactg
     4981 tcggcagagg catcttcaac gatggccttt cctttatcgc aatgatggca tttgtaggag
     5041 ccaccttcct tttccactat cttcacaata aagtgacaga tagctgggca atggaatccg
     5101 aggaggtttc cggatattac cctttgttga aaagtctcaa ttgccctttg gtcttctgag
     5161 actgtatctt tgatattttt ggagtagaca agtgtgtcgt gctccacatg ttgaccggtg
     5221 gcgcgccggc actggccgtc gttttacaac gaattcctgc agcccggggg atccactagt
     5281 tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat
     5341 tccgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5401 aattccacac aacatacgag ccggaaghca taaagtgtaa agcctggggt gcctaatgag
     5461 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5521 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5581 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5701 agaacatgaa ggccttgaca ggatatattg gcgggtaaac taagtcgctg tatgtgtttg
     5761 tttgagatct catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5821 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5881 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5941 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6001 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6061 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6121 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6181 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6241 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6301 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6361 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6421 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6481 ggattttggt catggttaca accaattaac caattctgat tagaaaaact catcgagcat
     6541 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     6601 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     6661 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     6721 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     6781 aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     6841 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     6901 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     6961 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     7021 tgttttccct gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     7081 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     7141 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     7201 cccatacaat ccatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     7261 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctggagcaag acgtttcccg
     7321 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     7381 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
     7441 gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc atcttcccga
     7501 caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt ccacctacaa
     7561 caaagctctc atcaaccgtg gctccctcac tttctggctg gatgatgggg cgattcaggc
     7621 gatccccatc caacagcccg ccgtcgagcg ggctttttta tccccggaag cctgtggata
     7681 gagggtagtt atccacgtga aaccgctaat gccccgcaaa gccttgattc acggggcttt
     7741 ccggcccgct ccaaaaacta tccacgtgaa atcgctaatc agggtacgtg aaatcgctaa
     7801 tcggagtacg tgaaatcgct aataaggtca cgtgaaatcg ctaatcaaaa aggcacgtga
     7861 gaacgctaat agccctttca gatcaacagc ttgcaaacac ccctcgctcc ggcaagtagt
     7921 tacagcaagt agtatgttca attagctttt caattatgaa tatatatatc aattattggt
     7981 cgcccttggc ttgtggacaa tgcgctacgc gcaccggctc cgcccgtgga caaccgcaag
     8041 cggttgccca ccgtcgagcg ccagcgcctt tgcccacaac ccggcggccg gccgcaacag
     8101 atcgttttat aaattttttt ttttgaaaaa gaaaaagccc gaaaggcggc aacctctcgg
     8161 gcttctggat ttccgatccc cggaattaga tcttggcagg atatattgtg gtgtaacgtt
     8221 atcagcttgc atgccggtcg atctagtaac atagatgaca ccgcgcgcga taatttatcc
     8281 tagtttgcgc gctatatttt gttttctatc gcgtattaaa tgtataattg cgggactcta
     8341 atcaaaaaac ccatctcata aataacgtca tgcattacat gttaattatt acatgcttaa
     8401 cgtaattcaa cagaaattat atgataatca tcgcaagacc ggcaacagga ttcaatctta
     8461 agaaacttta ttgccaaatg tttgaacgat ctgcttgact ctagctagag tccgaacccc
     8521 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     8581 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     8641 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     8701 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     8761 cctgggtcac gacgagatcc tcgccgtcgg gcatccgcgc cttgagcctg gcgaacagtt
     8821 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     8881 ccatccgagt acgtcctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     8941 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     9001 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     9061 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     9121 acgatagccg cgctgcctcg tcttggagtt cattcagggc accggacagg tcggtcttga
     9181 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     9241 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     9301 cgtgcaatcc atcttgttca atcatgcctc gatcgagttg agagtgaata tgagactcta
     9361 attggatacc gaggggaatt tatggaacgt cagtggagca tttttgacaa gaaatatttg
     9421 ctagctgata gtgaccttag gcgacttttg aacgcgcaat aatggtttct gacgtatgtg
     9481 cttagctcat taaactccag aaacccgcgg ctgagtggct ccttcaacgt tgcggttctg
     9541 tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc gggggtcata acgtgactcc
     9601 cttaattctc atgtatcgat aacattaacg tttacaattt cgcgccattc gccattcagg
     9661 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     9721 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     9781 cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga attgggtacc
     9841 gggccccccc tcgaggtcga cggtatcgat
//
