LOCUS       Exported                5141 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJH370, complete sequence.
ACCESSION   AF316553
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5141)
  AUTHORS   Hu JC, OShea EK, Kim PS, Sauer RT.
  TITLE     Sequence requirements for coiled-coils: analysis with lambda 
            repressor-GCN4 leucine zipper fusions
  JOURNAL   Science 250 (4986), 1400-1403 (1990)
  PUBMED    2147779
REFERENCE   2  (bases 1 to 5141)
  AUTHORS   Hu JC.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-OCT-2000) Biochemistry & Biophysics, Texas A&M 
            University, TAMU 2128, College Station, TX 77843-2128, USA
REFERENCE   3  (bases 1 to 5141)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5141
                     /organism="Cloning vector pJH370"
                     /mol_type="other DNA"
                     /db_xref="taxon:151159"
     regulatory      6..63
                     /regulatory_class="promoter"
                     /note="lacUV5"
     promoter        28..58
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     regulatory      28..34
                     /regulatory_class="minus_35_signal"
     regulatory      52..57
                     /regulatory_class="minus_10_signal"
     misc_binding    64..84
                     /label=lac operator binding site
                     /bound_moiety="lac operator"
     protein_bind    66..82
                     /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)."
     regulatory      91..94
                     /regulatory_class="ribosome_binding_site"
     CDS             103..603
                     /codon_start=1
                     /product="cI-GCN4 repressor fusion protein"
                     /label=cI-GCN4 repressor fusion protein
                     /protein_id="AAK07887.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTHMKQLEDKVEELLSKNYHLENEVARLKKLV
                     GER"
     CDS             529..585
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     regulatory      611..738
                     /regulatory_class="terminator"
                     /note="T-phi; T7 transcription terminator"
     terminator      670..717
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             2202..2393
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    2495..2635
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2821..3409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3520..4033
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(4075..4935)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="bla"
                     /protein_id="AAK07886.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4936..5040)
                     /gene="bla"
                     /label=AmpR promoter