LOCUS       Exported                6093 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pVMExp14367, complete sequence.
ACCESSION   MG599491
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6093)
  AUTHORS   Verma V, Kaur C, Grover P, Gupta A, Chaudhary VK.
  TITLE     Biotin-tagged proteins: Reagents for efficient ELISA-based 
            serodiagnosis and phage display-based affinity selection
  JOURNAL   PLoS ONE 13 (1), e0191315 (2018)
  PUBMED    29360877
REFERENCE   2  (bases 1 to 6093)
  AUTHORS   Chaudhary VK, Gupta A, Verma V, Kaur C, Grover P.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-DEC-2017) Centre for Innovation in Infectious Disease 
            Research, Education, and Training, University of Delhi South Campus,
            Benito Juarez Road, New Delhi, New Delhi 110021, India
REFERENCE   3  (bases 1 to 6093)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6093
                     /organism="Cloning vector pVMExp14367"
                     /mol_type="other DNA"
                     /db_xref="taxon:2079766"
     regulatory      1..44
                     /regulatory_class="promoter"
                     /note="T7 promoter lac operator"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    98..127
                     /label=deca-histidine tag
                     /note="deca-histidine tag"
     CDS             98..127
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     misc_feature    143..163
                     /label=TEV protease cleavage site
                     /note="TEV protease cleavage site"
     CDS             143..163
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    complement(169..179)
                     /label=BsaI-5 cloning site
                     /note="BsaI-5 cloning site"
     misc_feature    180..2083
                     /label=stuffer
                     /note="stuffer"
     promoter        180..625
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             626..2047
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /protein_id="AUW40200.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            626..2047
                     /gene="sacB"
                     /label=sacB
     misc_feature    2084..2094
                     /label=BsaI-3 cloning site
                     /note="BsaI-3 cloning site"
     misc_feature    2091..2117
                     /label=spacer
                     /note="spacer"
     misc_feature    2118..2162
                     /label=biotin acceptor peptide
                     /note="biotin acceptor peptide"
     CDS             2118..2162
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     regulatory      2230..2277
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      2230..2277
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2314..2769
                     /label=f1 origin
                     /note="f1 origin"
     promoter        2796..2900
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2901..3761
                     /codon_start=1
                     /product="beta lactamase"
                     /label=beta lactamase
                     /protein_id="AUW40201.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3932..4520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3932..4519
                     /note="ColE1 ori; deleted of rop gene"
     CDS             complement(4625..5707)
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI"
                     /label=lacI
                     /protein_id="AUW40202.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(4625..5707)
                     /gene="lacI"
                     /label=lacI
     promoter        complement(5708..5785)
                     /gene="lacI"
                     /label=lacI promoter