LOCUS       Exported               11498 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pDEST162, complete sequence.
ACCESSION   MH107059
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11498)
  AUTHORS   Wu FL, Liu Y, Zhang HN, Jiang HW, Cheng L, Guo SJ, Deng JY, Bi LJ, 
            Zhang XE, Gao HF, Tao SC.
  TITLE     Global profiling of PknG interactions using a human proteome 
            microarray reveals novel connections with CypA
  JOURNAL   Proteomics, e1800265 (2018) In press
  PUBMED    30281201
REFERENCE   2  (bases 1 to 11498)
  AUTHORS   Wu FL, Tao SC.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAR-2018) Shanghai Center for Systems Biomedicine, 
            Shanghai Jiao Tong University, Dongchuan Road, No. 800, Shanghai, 
            Shanghai 200210, China
REFERENCE   3  (bases 1 to 11498)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..11498
                     /organism="Expression vector pDEST162"
                     /mol_type="other DNA"
                     /db_xref="taxon:2482960"
     rep_origin      475..2264
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     CDS             complement(2570..4495)
                     /codon_start=1
                     /product="Epstein-Barr nuclear antigen 1, also known as 
                     EBNA-1"
                     /label=EBNA1
                     /note="crucial for latent viral infection, and for episomal
                     amplification of vectors containing the oriP origin (Okita 
                     et al., 2013)"
                     /translation="MSDEGPGTGPGNGLGEKGDTSGPEGSGGSGPQRRGGDNHGRGRGR
                     GRGRGGGRPGAPGGSGSGPRHRDGVRRPQKRPSCIGCKGTHGGTGAGAGAGGAGAGGAG
                     AGGGAGAGGGAGGAGGAGGAGAGGGAGAGGGAGGAGGAGAGGGAGAGGGAGGAGAGGGA
                     GGAGGAGAGGGAGAGGGAGGAGAGGGAGGAGGAGAGGGAGAGGAGGAGGAGAGGAGAGG
                     GAGGAGGAGAGGAGAGGAGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGGAGAG
                     GGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGAGGAGAGGGGRGRGGSGGRGRG
                     GSGGRGRGGSGGRRGRGRERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPP
                     PGRRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGR
                     RKKGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTDEGTWVAGVFVYGGSKTSL
                     YNLRRGTALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLK
                     DAIKDLVMTKPAPTCNIRVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDE
                     GEEGQE"
     regulatory      5016..5120
                     /regulatory_class="promoter"
     promoter        5016..5120
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5121..5981
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /protein_id="AYN74339.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6152..6740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      6968..7265
                     /regulatory_class="promoter"
     promoter        6968..7265
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             7343..7942
                     /codon_start=1
                     /product="puromycin N-acetyltransferase"
                     /label=puromycin N-acetyltransferase
                     /note="confers resistance to puromycin"
                     /protein_id="AYN74340.1"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    8033..8167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      8426..8805
                     /regulatory_class="enhancer"
     enhancer        8426..8805
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      8806..9009
                     /regulatory_class="promoter"
     promoter        8806..9009
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             9181..9204
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             9241..9264
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             9271..9288
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             9295..9312
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     protein_bind    9325..9449
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      9474..9504
                     /regulatory_class="promoter"
     promoter        9474..9504
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             9558..10217
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=chloramphenicol acetyltransferase
                     /note="confers resistance to chloramphenicol"
                     /protein_id="AYN74341.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             10559..10864
                     /codon_start=1
                     /product="CcdB"
                     /label=CcdB
                     /note="bacterial toxin that poisons DNA gyrase"
                     /protein_id="AYN74342.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(10905..11029)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             11037..11054
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             11061..11084
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    11089..11223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"