LOCUS       Exported                4115 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pAK-TAG-T, complete sequence.
ACCESSION   KX147099
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4115)
  AUTHORS   Luo D, Wen C, Zhao R, Liu X, Liu X, Cui J, Liang JG, Liang P.
  TITLE     High Level Expression and Purification of Recombinant Proteins from 
            Escherichia coli with AK-TAG
  JOURNAL   PLoS ONE 11 (5), E0156106 (2016)
  PUBMED    27214237
REFERENCE   2  (bases 1 to 4115)
  AUTHORS   Luo D.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-APR-2016) Department of Biochemistry & Molecular 
            Biology, School of Life Sciences, Sichuan University, No. 24 South 
            Section 1, Yihuan Road, Cheng Du, Si Chuan 610064, China
REFERENCE   3  (bases 1 to 4115)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4115
                     /organism="Expression vector pAK-TAG-T"
                     /mol_type="other DNA"
                     /db_xref="taxon:1853219"
     regulatory      10..54
                     /regulatory_class="promoter"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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)."
     protein_bind    62..78
                     /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             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    145..798
                     /note="similar to adenylate kinase; AK"
     misc_feature    799..816
                     /label=thrombin cleavage site
                     /note="thrombin cleavage site"
     CDS             799..816
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     misc_feature    817..846
                     /label=multiple cloning site
                     /note="multiple cloning site"
     terminator      862..956
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1000..1659
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      1724..1810
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    1965..2105
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2291..2879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3050..3910)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3911..4015)
                     /gene="bla"
                     /label=AmpR promoter