LOCUS       Exported                6640 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pGWnV4 DNA, complete sequence.
ACCESSION   LC030200
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6640)
  AUTHORS   Nishimura K, Matsunami E, Yoshida S, Kohata S, Yamauchi J, Jisaka M,
            Nagaya T, Yokota K, Nakagawa T.
  TITLE     The tyrosine-sorting motif of the vacuolar sorting receptor VSR4 
            from Arabidopsis thaliana, which is involved in the interaction 
            between VSR4 and AP1M2, mu1-adaptin type 2 of clathrin adaptor 
            complex 1 subunits, participates in the post-Golgi sorting of VSR4
  JOURNAL   Biosci. Biotechnol. Biochem. 80 (4), 694-705 (2016)
  PUBMED    26745465
REFERENCE   2  (bases 1 to 6640)
  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 6640)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     constructed using pUC119.
FEATURES             Location/Qualifiers
     source          1..6640
                     /organism="Gateway vector pGWnV4"
                     /mol_type="other DNA"
                     /db_xref="taxon:1619210"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /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)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      276..1109
                     /regulatory_class="promoter"
                     /note="35S promoter"
     promoter        763..1108
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    complement(1130..1254)
                     /label=attR1
                     /note="attR1"
     protein_bind    1130..1254
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1291..1321
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1375..2034
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="BAU29945.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            1375..2034
                     /gene="cat"
                     /label=cat
     CDS             2376..2681
                     /codon_start=1
                     /gene="ccdB"
                     /label=ccdB
                     /protein_id="BAU29946.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            2376..2681
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    2722..2846
                     /label=attR2 (replacement of t with c at 2846)
                     /note="attR2 (replacement of t with c at 2846)"
     protein_bind    complement(2722..2846)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2857..3486
                     /note="n-terminal fragment of yellow fluorescent protein 
                     (replacement of gcc with aag at 3475-3477)"
     CDS             2857..3375
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     regulatory      3492..3768
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     terminator      3508..3760
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3769..3785)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3998..4453
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4735..4839
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4840..5700
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase"
                     /label=bla
                     /protein_id="BAU29947.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4840..5700
                     /gene="bla"
                     /label=bla
     rep_origin      complement(5861..6461)
                     /direction=LEFT
                     /label=ColE1 Ori
                     /note="ColE1 Ori"
     rep_origin      5871..6459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"