LOCUS       Exported                5451 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pETAT-2, complete sequence.
ACCESSION   AF525442
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5451)
  AUTHORS   Yang Y, Ma J, Song Z, Wu M.
  TITLE     HIV-1 TAT-mediated protein transduction and subcellular localization
            using novel expression vectors
  JOURNAL   FEBS Lett. 532 (1-2), 36-44 (2002)
  PUBMED    12459459
REFERENCE   2  (bases 1 to 5451)
  AUTHORS   Yang YH, Wu M.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUN-2002) Department of Molecular and Cellular 
            Biology, School of Life Sciences, University of Science and 
            Technology of China, 96 Jingzai Road, Hefei, Anhui 230026, P.R. 
            China
REFERENCE   3  (bases 1 to 5451)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5451
                     /organism="Expression vector pETAT-2"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:206384"
     terminator      26..73
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      complement(26..72)
                     /regulatory_class="terminator"
                     /note="T7"
     misc_feature    complement(140..157)
                     /label=histidine tag
                     /note="histidine tag"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    158..211
                     /note="multiple cloning site; MCS"
     misc_feature    complement(212..244)
                     /note="contains mutation; Region: synthetic HIV-1 TAT 
                     protein transduction domain (PTD)"
     RBS             255..277
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    292..316
                     /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(317..335)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(319..335)
                     /regulatory_class="promoter"
                     /note="T7"
     promoter        644..721
                     /gene="lacI"
                     /label=lacI promoter
     CDS             722..1804
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI"
                     /label=lacI
                     /protein_id="AAO20027.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            722..1804
                     /gene="lacI"
                     /label=lacI
     CDS             2613..2804
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    2906..3048
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3234..3822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3235
                     /label=pBR322
                     /note="pBR322"
     CDS             complement(3993..4853)
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase"
                     /label=bla
                     /protein_id="AAO20028.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3993..4853)
                     /gene="bla"
                     /label=bla
     promoter        complement(4854..4958)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4985..5440)
                     /direction=LEFT
                     /label=f1
                     /note="f1"