LOCUS       40924_13860        6081 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Gateway vector pcVGW DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6081)
  AUTHORS   Nishimura K, Ishikawa S, Matsunami E, Yamauchi J, Homma K, Faulkner 
            C, Oparka K, Jisaka M, Nagaya T, Yokota K, Nakagawa T.
  TITLE     New Gateway-compatible vectors for a high-throughput protein-protein
            interaction analysis by a bimolecular fluorescence complementation 
            (BiFC) assay in plants and their application to a plant clathrin 
            structure analysis
  JOURNAL   Biosci. Biotechnol. Biochem. 79 (12), 1995-2006 (2015)
  PUBMED    26193449
REFERENCE   2  (bases 1 to 6081)
  AUTHORS   Nishimura K, Matsunami E.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-FEB-2015) Contact:Kohji Nishimura Shimane University, 
            Organization of Research; 1060 Nishikawatsu, Matsue, Shimane 
            690-8504, Japan
REFERENCE   3  (bases 1 to 6081)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6081)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biosci. 
            Biotechnol. Biochem."; date: "2015"; volume: "79"; issue: "12"; 
            pages: "1995-2006"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-FEB-2015) Contact:Kohji Nishimura Shimane University, 
            Organization of Research; 1060 Nishikawatsu, Matsue, Shimane 
            690-8504, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     constructed using pUC119.
FEATURES             Location/Qualifiers
     source          1..6081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        745..1090
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    1111..1197
                     /label=c-terminal fragment of yellow fluorescent protein
                     /note="c-terminal fragment of yellow fluorescent protein"
     protein_bind    1207..1331
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1368..1398
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1452..2108
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2453..2755
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2799..2923)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      2949..3201
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3210..3226)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3439..3894
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4176..4280
                     /label=AmpR promoter
     CDS             4281..5138
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5312..5900
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 catgcctgca ggtccccaga ttagcctttt caatttcaga aagaatgcta acccacagat
      301 ggttagagag gcttacgcag caggtctcat caagacgatc tacccgagca ataatctcca
      361 ggaaatcaaa taccttccca agaaggttaa agatgcagtc aaaagattca ggactaactg
      421 catcaagaac acagagaaag atatatttct caagatcaga agtactattc cagtatggac
      481 gattcaaggc ttgcttcaca aaccaaggca agtaatagag attggagtct ctaaaaaggt
      541 agttcccact gaatcaaagg ccatggagtc aaagattcaa atagaggacc taacagaact
      601 cgccgtaaag actggcgaac agttcataca gagtctctta cgactcaatg acaagaagaa
      661 aatcttcgtc aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac
      721 agtctcagaa gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct
      781 cctcggattc cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg
      841 tggctcctac aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc
      901 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt
      961 tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga
     1021 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
     1081 ggagagaaca cgggggactc tagaatgagc gaccccaacg agaagcgcga tcacatggtc
     1141 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggct
     1201 gttatcacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat
     1261 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
     1321 cagtcactat ggcggcccct cgaggggccg cattaggcac cccaggcttt acactttatg
     1381 cttccggctc gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta
     1441 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     1501 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     1561 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     1621 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa
     1681 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     1741 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     1801 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     1861 ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg
     1921 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     1981 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     2041 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     2101 gcggggcgta aacgcgtgga tccggcttac taaaagccag ataacagtat gcgtatttgc
     2161 gcgctgattt ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa
     2221 gaggtatgct atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct
     2281 caaggcatat atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc
     2341 cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc
     2401 cggtttattg aaatgaacgg ctcttttgct gacgagaaca ggggctggtg aaatgcagtt
     2461 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
     2521 tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
     2581 agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
     2641 gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct
     2701 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat
     2761 gtcaggctcc cttatacaca gccagtctgc aggtcgacca tagtgactgg atatgttgtg
     2821 ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta
     2881 tatcatttta cgtttctcgt tcagctttct tgtacaaagt ggtgataacc tagagctcga
     2941 atttccccga tcgttcaaac atttggcaat aaagtttctt aagattgaat cctgttgccg
     3001 gtcttgcgat gattatcata taatttctgt tgaattacgt taagcatgta ataattaaca
     3061 tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg caattataca
     3121 tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg
     3181 tgtcatctat gttactagat cgggaattca ctggccgtcg ttttacaacg tcgtgactgg
     3241 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     3301 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     3361 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3421 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3481 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3541 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3601 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     3661 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3721 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3781 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3841 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     3901 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     3961 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4021 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4081 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4141 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4201 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4261 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4321 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4381 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4441 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4501 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4561 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4621 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4681 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4741 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4801 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4861 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4921 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4981 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5041 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5101 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5161 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5221 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5281 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5341 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5401 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     5461 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5521 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5581 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5641 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5701 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5761 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5821 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5881 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5941 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6001 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6061 cagtgagcga ggaagcggaa g
//
