LOCUS       40924_16275        8643 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Plant expression vector pDuExB2 (pDuExD7), complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8643)
  AUTHORS   Fujikawa Y, Kato N.
  TITLE     Split luciferase complementation assay to study protein-protein 
            interactions in Arabidopsis protoplasts
  JOURNAL   Plant J. 52 (1), 185-195 (2007)
  PUBMED    17662028
REFERENCE   2  (bases 1 to 8643)
  AUTHORS   Kato N.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-APR-2007) Dapartment of Biological Sciences, Louisiana
            State University, 226 Life Sciences Building, Baton Rouge, LA 
            70803-1715, USA
REFERENCE   3  (bases 1 to 8643)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8643)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant J."; 
            date: "2007"; volume: "52"; issue: "1"; pages: "185-195"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-APR-2007) Dapartment of Biological Sciences, Louisiana State 
            University, 226 Life Sciences Building, Baton Rouge, LA 70803-1715, 
            USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             36..281
                     /codon_start=1
                     /product="ChRluc"
                     /label=ChRluc
                     /note="C-terminal fragment of the Renilla luciferase
                     protein"
                     /protein_id="ABU49628.1"
                     /translation="KPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFP
                     NTEFVKVKGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             282..299
                     /codon_start=1
                     /label=tetracysteine tag
                     /note="tetracysteine peptide that binds biarsenical
                     labeling reagents"
                     /translation="CCPGCC"
     terminator      324..576
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(585..601)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(909..1565)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     misc_recomb     1594..1627
                     /label=loxP
                     /note="loxP"
     protein_bind    1594..1627
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        1642..2087
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             2088..3506
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     promoter        3577..3681
                     /label=AmpR promoter
     CDS             3682..4539
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4713..5301
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5418..5434
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5442..5460
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(5526..5559)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        6079..6424
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    6436..6491
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             6508..6753
                     /codon_start=1
                     /product="ChRluc"
                     /label=ChRluc
                     /note="C-terminal fragment of the Renilla luciferase
                     protein"
                     /protein_id="ABU49629.1"
                     /translation="KPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFP
                     NTEFVKVKGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     protein_bind    6790..6914
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        7061..7163
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             7164..7820
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             8165..8467
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(8511..8635)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ctagaggtgg tggcggttca ggcggaggtg gctctaagcc cgacgtcgtc cagattgtcc
       61 gcaactacaa cgcctacctt cgggccagcg acgatctgcc taagatgttc atcgagtccg
      121 accctgggtt cttttccaac gctattgtcg agggagctaa gaagttccct aacaccgagt
      181 tcgtgaaggt gaagggcctc cacttcagcc aggaggacgc tccagatgaa atgggtaagt
      241 acatcaagag cttcgtggag cgcgtgctga agaacgagca gtgctgcccc gggtgctgct
      301 aagagctcag ctcgaatttc cccgatcgtt caaacatttg gcaataaagt ttcttaagat
      361 tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc
      421 atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag
      481 tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata
      541 aattatcgcg cgcggtgtca tctatgttac tagatcggga attcactggc cgtcgtttta
      601 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
      661 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
      721 cgcagcctga atggcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
      781 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
      841 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg cagctcgata
      901 tcaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg
      961 ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc
     1021 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
     1081 ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac
     1141 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
     1201 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa
     1261 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
     1321 agctcaccgt ctttcattgc catacgaaat tccggatgag cattcatcag gcgggcaaga
     1381 atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc
     1441 gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca
     1501 aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc
     1561 tccattttag cttccttagc tcctgaaaga tccataactt cgtatagcat acattatacg
     1621 aagttatgcg gccgcgacgt ccacatatac ctgccgttca ctattattta gtgaaatgag
     1681 atattatgat attttctgaa ttgtgattaa aaaggcaact ttatgcccat gcaacagaaa
     1741 ctataaaaaa tacagagaat gaaaagaaac agatagattt tttagttctt taggcccgta
     1801 gtctgcaaat ccttttatga ttttctatca aacaaaagag gaaaatagac cagttgcaat
     1861 ccaaacgaga gtctaataga atgaggtcga aaagtaaatc gcgcgggttt gttactgata
     1921 aagcaggcaa gacctaaaat gtgtaaaggg caaagtgtat actttggcgt caccccttac
     1981 atattttagg tcttttttta ttgtgcgtaa ctaacttgcc atcttcaaac aggagggctg
     2041 gaagaagcag accgctaaca cagtacataa aaaaggagac atgaacgatg aacatcaaaa
     2101 agtttgcaaa acaagcaaca gtattaacct ttactaccgc actgctggca ggaggcgcaa
     2161 ctcaagcgtt tgcgaaagaa acgaaccaaa agccatataa ggaaacatac ggcatttccc
     2221 atattacacg ccatgatatg ctgcaaatcc ctgaacagca aaaaaatgaa aaatatcaag
     2281 ttcctgagtt cgattcgtcc acaattaaaa atatctcttc tgcaaaaggc ctggacgttt
     2341 gggacagctg gccattacaa aacgctgacg gcactgtcgc aaactatcac ggctaccaca
     2401 tcgtctttgc attagccgga gatcctaaaa atgcggatga cacatcgatt tacatgttct
     2461 atcaaaaagt cggcgaaact tctattgaca gctggaaaaa cgctggccgc gtctttaaag
     2521 acagcgacaa attcgatgca aatgattcta tcctaaaaga ccaaacacaa gaatggtcag
     2581 gttcagccac atttacatct gacggaaaaa tccgtttatt ctacactgat ttctccggta
     2641 aacattacgg caaacaaaca ctgacaactg cacaagttaa cgtatcagca tcagacagct
     2701 ctttgaacat caacggtgta gaggattata aatcaatctt tgacggtgac ggaaaaacgt
     2761 atcaaaatgt acagcagttc atcgatgaag gcaactacag ctcaggcgac aaccatacgc
     2821 tgagagatcc tcactacgta gaagataaag gccacaaata cttagtattt gaagcaaaca
     2881 ctggaactga agatggctac caaggcgaag aatctttatt taacaaagca tactatggca
     2941 aaagcacatc attcttccgt caagaaagtc aaaaacttct gcaaagcgat aaaaaacgca
     3001 cggctgagtt agcaaacggc gctctcggta tgattgagct aaacgatgat tacacactga
     3061 aaaaagtgat gaaaccgctg attgcatcta acacagtaac agatgaaatt gaacgcgcga
     3121 acgtctttaa aatgaacggc aaatggtacc tgttcactga ctcccgcgga tcaaaaatga
     3181 cgattgacgg cattacgtct aacgatattt acatgcttgg ttatgtttct aattctttaa
     3241 ctggcccata caagccgctg aacaaaactg gccttgtgtt aaaaatggat cttgatccta
     3301 acgatgtaac ctttacttac tcacacttcg ctgtacctca agcgaaagga aacaatgtcg
     3361 tgattacaag ctatatgaca aacagaggat tctacgcaga caaacaatca acgtttgcgc
     3421 ctagcttcct gctgaacatc aaaggcaaga aaacatctgt tgtcaaagac agcatccttg
     3481 aacaaggaca attaacagtt aacaaataaa aacgcaaaag aaaatgccga tatcctattg
     3541 gcattgacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3601 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3661 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3721 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3781 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3841 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3901 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3961 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4021 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4081 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4141 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4201 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4261 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4321 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4381 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4441 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4501 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4561 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4621 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4681 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4741 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4801 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4861 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4921 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4981 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5041 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5101 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5161 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5221 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5281 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5341 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5401 attaccgcct tacgcgtgta aaacgacggc cagtagatct gtaatacgac tcactatagg
     5461 gcgctagctg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5521 gccgcataac ttcgtatagc atacattata cgaagttatc agtcgacaag cttgcatgcc
     5581 tgcaggtccc cagattagcc ttttcaattt cagaaagaat gctaacccac agatggttag
     5641 agaggcttac gcagcaggtc tcatcaagac gatctacccg agcaataatc tccaggaaat
     5701 caaatacctt cccaagaagg ttaaagatgc agtcaaaaga ttcaggacta actgcatcaa
     5761 gaacacagag aaagatatat ttctcaagat cagaagtact attccagtat ggacgattca
     5821 aggcttgctt cacaaaccaa ggcaagtaat agagattgga gtctctaaaa aggtagttcc
     5881 cactgaatca aaggccatgg agtcaaagat tcaaatagag gacctaacag aactcgccgt
     5941 aaagactggc gaacagttca tacagagtct cttacgactc aatgacaaga agaaaatctt
     6001 cgtcaacatg gtggagcacg acacacttgt ctactccaaa aatatcaaag atacagtctc
     6061 agaagaccaa agggcaattg agacttttca acaaagggta atatccggaa acctcctcgg
     6121 attccattgc ccagctatct gtcactttat tgtgaagata gtggaaaagg aaggtggctc
     6181 ctacaaatgc catcattgcg ataaaggaaa ggccatcgtt gaagatgcct ctgccgacag
     6241 tggtcccaaa gatggacccc cacccacgag gagcatcgtg gaaaaagaag acgttccaac
     6301 cacgtcttca aagcaagtgg attgatgtga tatctccact gacgtaaggg atgacgcaca
     6361 atcccactat ccttcgcaag acccttcctc tatataagga agttcatttc atttggagag
     6421 aacagtattt ttacaacaat taccaacaac aacaaacaac aaacaacatt acaattacta
     6481 tttacaatta caggatctct cgagatgaag cccgacgtcg tccagattgt ccgcaactac
     6541 aacgcctacc ttcgggccag cgacgatctg cctaagatgt tcatcgagtc cgaccctggg
     6601 ttcttttcca acgctattgt cgagggagct aagaagttcc ctaacaccga gttcgtgaag
     6661 gtgaagggcc tccacttcag ccaggaggac gctccagatg aaatgggtaa gtacatcaag
     6721 agcttcgtgg agcgcgtgct gaagaacgag cagggtggtg gcggttcagg cggaggtggc
     6781 tctgtcgaga caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc
     6841 aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat
     6901 atccagtcac tatggcggcc gctaagttgg cagcatcacc cgacgcactt tgcgccgaat
     6961 aaatacctgt gacggaagat cacttcgcag aataaataaa tcctggtgtc cctgttgata
     7021 ccgggaagcc ctgggccaac ttttggcgaa aatgagacgt tgatcggcac gtaagaggtt
     7081 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat
     7141 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     7201 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     7261 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     7321 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     7381 tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     7441 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     7501 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     7561 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
     7621 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     7681 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     7741 atgccgtctg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     7801 atgagtggca gggcggggcg taaacgcgtg gatccggctt actaaaagcc agataacagt
     7861 atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa
     7921 gtatgtcaaa aagaggtgtg ctatgaagca gcgtattaca gtgacagttg acagcgacag
     7981 ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg
     8041 cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg
     8101 gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa cagggactgg
     8161 tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga
     8221 tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc
     8281 acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga
     8341 aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga
     8401 agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg
     8461 gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgac catagtgact
     8521 ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata
     8581 tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa gtggtcagat
     8641 cct
//
