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NX32_GW vector (V009827#)

Basic Vector Information

Basic Vector Information
Vector Name NX32_GW Antibiotic Resistance Ampicillin
Length 8899 bp Type Expression vector
Source Lalonde S, Sero A, Pratelli R, Pilot G, Chen J, Sardi MI, Parsa SA, Kim DY, Acharya BR, Stein EV, Hu HC, Villiers F, Takeda K, Yang Y, Han YS, Schwacke R, Chiang W, Kato N, Loque D, Assmann SM, Kwak J

NX32_GW vector Vector Map

2-micron TRP1 promoter F1 origin M13 fwd ADH1 promoter NubG Gateway recombination cassette containing the attR sites, ccdB and CamR genes 3x HA-tag stop codon ADH1 M13 rev lac operator lac promoter ColE1 AmpR AmpR promoter

NX32_GW vector Sequence

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LOCUS       Exported                8899 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector NX32_GW, complete sequence.
ACCESSION   HQ700952
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8899)
  AUTHORS   Lalonde S, Sero A, Pratelli R, Pilot G, Chen J, Sardi MI, Parsa SA, 
            Kim DY, Acharya BR, Stein EV, Hu HC, Villiers F, Takeda K, Yang Y, 
            Han YS, Schwacke R, Chiang W, Kato N, Loque D, Assmann SM, Kwak JM, 
            Schroeder JI, Rhee SY, Frommer WB.
  TITLE     A Membrane Protein/Signaling Protein Interaction Network for 
            Arabidopsis Version AMPv2
  JOURNAL   Front Physiol 1, 24 (2010)
  PUBMED    21423366
REFERENCE   2  (bases 1 to 8899)
  AUTHORS   Lalonde S, Frommer WB.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-DEC-2010) Plant Biology, Carnegie Institution, 260 
            Panama St, Stanford, CA 94305, USA
REFERENCE   3  (bases 1 to 8899)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8899
                     /organism="Expression vector NX32_GW"
                     /mol_type="other DNA"
                     /db_xref="taxon:1027257"
     rep_origin      2..1348
                     /label=2-micron
                     /note="2-micron"
     rep_origin      2..1344
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(965..1012)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        1603..1883
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     gene            1782..2604
                     /gene="TRP1"
                     /label=TRP1
     regulatory      1782..1883
                     /regulatory_class="promoter"
                     /gene="TRP1"
     CDS             1884..2558
                     /codon_start=1
                     /gene="TRP1"
                     /product="TRP1"
                     /label=TRP1
                     /protein_id="AED88266.1"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     regulatory      2559..2604
                     /regulatory_class="terminator"
                     /gene="TRP1"
     rep_origin      complement(2661..3119)
                     /direction=LEFT
                     /label=F1 origin
                     /note="F1 origin"
     rep_origin      complement(2661..3116)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     3261..3277
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3605..4006
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     regulatory      3650..4010
                     /regulatory_class="promoter"
                     /note="ADH1"
     misc_feature    4011..4121
                     /note="NubG; N-terminal moiety of ubiquitin (ubi4)"
     misc_difference 4047..4049
                     /label=mutation Ile to Gly
                     /note="mutation Ile to Gly"
     misc_feature    4131..6398
                     /note="Gateway recombination cassette containing the attR 
                     sites, ccdB and CamR genes"
     misc_recomb     4131..4255
                     /label=attR1
                     /note="attR1"
     protein_bind    4131..4255
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4684..4989)
                     /codon_start=1
                     /product="ccdB"
                     /label=ccdB
                     /protein_id="AED88267.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(5334..5993)
                     /codon_start=1
                     /product="CamR"
                     /label=CamR
                     /note="chloramphenicol acetyl transferase"
                     /protein_id="AED88268.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(5994..6096)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     misc_recomb     6274..6398
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(6274..6398)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    6412..6501
                     /label=3x HA-tag
                     /note="3x HA-tag"
     CDS             6412..6501
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_feature    6511..6513
                     /label=stop codon
                     /note="stop codon"
     regulatory      6534..6724
                     /regulatory_class="terminator"
                     /note="ADH1"
     terminator      6534..6721
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6748..6764)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6772..6788
                     /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)."
     promoter        complement(6796..6826)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      7132..7747
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(7150..7738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7909..8769)
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /note="beta-lactamase"
                     /protein_id="AED88269.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8770..8874)
                     /gene="bla"
                     /label=AmpR promoter

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