LOCUS       Exported                5803 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pET15b, complete sequence.
ACCESSION   KX765180
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5803)
  AUTHORS   Villarreal FD, Chavez M, Ding Y, Fan J, Pan T, Tan C.
  TITLE     Synthetic microbial consortia enable rapid assembly of multi-protein
            complexes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5803)
  AUTHORS   Villarreal FD, Chavez M, Ding Y, Fan J, Pan T, Tan C.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-AUG-2016) Biomedical Engineering, University of 
            California, Davis, One Shields Ave, Davis, CA 95616, USA
REFERENCE   3  (bases 1 to 5803)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5803
                     /organism="Cloning vector pET15b"
                     /mol_type="other DNA"
                     /db_xref="taxon:1982515"
     regulatory      3..46
                     /regulatory_class="promoter"
     promoter        3..21
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    22..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)."
     RBS             72..80
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             86..445
                     /codon_start=1
                     /product="chagasin-FLAG"
                     /label=chagasin-FLAG
                     /note="protease inhibitor from Trypanozoma cruzi"
                     /protein_id="ARM20006.1"
                     /translation="MSHKVTKAHNGATLTVAVGELVEIQLPSNPTTGFAWYFEGGTKES
                     PNESMFTVENKYFPPDSKLLGAAGGTEHFHVTVKAAGTHAVNLTYMRPWTGPSHDSERF
                     TVYLKANDYKDDDDK"
     CDS             419..442
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      446..573
                     /regulatory_class="terminator"
     terminator      521..568
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        680..784
                     /gene="bla"
                     /label=AmpR promoter
     CDS             785..1645
                     /codon_start=1
                     /product="BlaI"
                     /label=BlaI
                     /note="ampicillin resistance protein"
                     /protein_id="ARM20007.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1816..2404
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2590..2732
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2834..3025)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4337..5419)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5420..5497)
                     /gene="lacI"
                     /label=lacI promoter