LOCUS       Exported               10755 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pMetYC_GW, complete sequence.
ACCESSION   HQ646605
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10755)
  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 10755)
  AUTHORS   Lalonde S, Frommer WB.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-NOV-2010) Plant Biology, Carnegie Institution, 260 
            Panama St, Stanford, CA 94305, USA
REFERENCE   3  (bases 1 to 10755)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10755
                     /organism="Expression vector pMetYC_GW"
                     /mol_type="other DNA"
                     /db_xref="taxon:1027262"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        241..259
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      277..659
                     /regulatory_class="promoter"
                     /note="Met25"
     promoter        309..659
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     misc_feature    660..2927
                     /note="Gateway recombination cassette containing the attR 
                     sites, ccdB and CamR genes"
     misc_recomb     660..784
                     /label=attR1
                     /note="attR1"
     protein_bind    660..784
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1213..1518)
                     /codon_start=1
                     /product="ccdB"
                     /label=ccdB
                     /protein_id="AED88257.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(1863..2522)
                     /codon_start=1
                     /product="CamR"
                     /label=CamR
                     /note="chloramphenicol acetyl transferase"
                     /protein_id="AED88256.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(2523..2625)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     misc_recomb     2803..2927
                     /label=attR1
                     /note="attR1"
     protein_bind    complement(2803..2927)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2929..2958
                     /label=linker
                     /note="linker"
     misc_feature    2959..3087
                     /note="Cub; C-terminal of ubiquitin (ubi4); start codon 
                     provided by fusion protein; stop codon at the end of VP16"
     misc_feature    3118..4365
                     /note="PLV; ProA-LexA-VP16; artificial transcription 
                     factor"
     misc_feature    3118..3495
                     /note="ProA; protease A; INSDC accession X96612"
     CDS             3139..3312
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             3316..3486
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     misc_feature    3505..4110
                     /note="lexA binding protein; INSDC accession J01643"
     CDS             3505..4110
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5-CTGTATATATATACAG-3."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     misc_feature    4128..4362
                     /note="VP16 DNA binding domain; UniProt accession P04486"
     CDS             4129..4362
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     misc_feature    4363..4365
                     /label=stop codon
                     /note="stop codon"
     polyA_signal    4558..4606
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        complement(4908..4926)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4933..4949)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5698..5808
                     /label=CEN
                     /note="CEN"
     misc_feature    5708..6870
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      6033..6870
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     gene            complement(7130..8745)
                     /gene="Leu2"
                     /label=Leu2
     regulatory      complement(7130..7231)
                     /regulatory_class="terminator"
                     /gene="Leu2"
     CDS             complement(7232..8326)
                     /codon_start=1
                     /gene="Leu2"
                     /product="Leu2"
                     /label=Leu2
                     /protein_id="AED88258.1"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(8339..8743)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     regulatory      complement(8340..8745)
                     /regulatory_class="promoter"
                     /gene="Leu2"
     promoter        8849..8953
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8954..9814
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /note="beta-lactamase"
                     /protein_id="AED88255.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            8954..9814
                     /gene="ampR"
                     /label=ampR
     rep_origin      9976..10591
                     /label=ColE1
                     /note="ColE1"
     rep_origin      9985..10573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"