LOCUS       Exported                4712 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLZ1, complete sequence.
ACCESSION   KF918333
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4712)
  AUTHORS   Zhu L, Klenner C, Kuhn A, Dalbey RE.
  TITLE     Both YidC and SecYEG are required for translocation of the 
            periplasmic loops 1 and 2 of the multispanning membrane protein TatC
  JOURNAL   J. Mol. Biol. 424 (5), 354-367 (2012)
  PUBMED    23058713
REFERENCE   2  (bases 1 to 4712)
  AUTHORS   Zhu L, Wasey A, White SH, Dalbey RE.
  TITLE     Charge composition features of model single-span membrane proteins 
            that determine selection of YidC and SecYEG translocase pathways in 
            Escherichia coli
  JOURNAL   J. Biol. Chem. 288 (11), 7704-7716 (2013)
  PUBMED    23355473
REFERENCE   3  (bases 1 to 4712)
  AUTHORS   Zhu L, Kaback HR, Dalbey RE.
  TITLE     YidC protein, a molecular chaperone for LacY protein folding via the
            SecYEG protein machinery
  JOURNAL   J. Biol. Chem. 288 (39), 28180-28194 (2013)
  PUBMED    23928306
REFERENCE   4  (bases 1 to 4712)
  AUTHORS   Soman R, Yuan J, Kuhn A, Dalbey RE.
  TITLE     The polarity and charge of the periplasmic loop determines the YidC 
            and Sec translocase requirement for the M13 procoat lep protein
  JOURNAL   J. Biol. Chem. (2013) In press
  PUBMED    24275657
REFERENCE   5  (bases 1 to 4712)
  AUTHORS   Zhu L, Dalbey R.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-NOV-2013) Department of Chemistry and Biochemistry, 
            The Ohio State University, RM580, W. 12th Ave., Columbus, OH 43210, 
            USA
REFERENCE   6  (bases 1 to 4712)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4712
                     /organism="Cloning vector pLZ1"
                     /mol_type="other DNA"
                     /note="High-copy and dual-promoter expression (or 
                     co-expression) vector system, compatible with ColE1, p15A, 
                     etc replicon vectors"
                     /db_xref="taxon:1441631"
     regulatory      20..38
                     /regulatory_class="promoter"
                     /note="T7 promoter; not functional for T7 RNA polymerase 
                     null strain; IPTG inducible promoter"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    38..332
                     /note="lac operator and LacUV5; compatible with T7 RNA 
                     polymerase null strain"
     protein_bind    39..63
                     /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        197..227
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    235..251
                     /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)."
     CDS             335..607
                     /codon_start=1
                     /product="LacZ"
                     /label=LacZ
                     /note="beta-galactosidase, alpha peptide; derived from INSD
                     CAA07592.1"
                     /protein_id="AHE74076.1"
                     /translation="MGDPKLLLEVPHAISSSPGNSLAVVLQRRDWENPGVTQLNRLAAH
                     PPFASWRNSEEARTDRPSQQLRSLIRLLTKPERKLSWLLPPLSNN"
     primer_bind     complement(397..413)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      545..676
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      604..651
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      704..790
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      707..750
                     /regulatory_class="terminator"
                     /note="rrnB_T1 terminator"
     regulatory      882..909
                     /regulatory_class="terminator"
                     /note="rrnB_T2 terminator"
     terminator      882..909
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      1006..1173
                     /regulatory_class="terminator"
                     /note="fd terminator"
     terminator      1009..1057
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     terminator      1119..1167
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             1313..1357
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      1409..1456
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1686..2501
                     /note="RSF origin; copy number above 100 per cell"
     CDS             complement(1686..2501)
                     /codon_start=1
                     /product="kanamicin resistance factor"
                     /label=kanamicin resistance factor
                     /note="derived from INSD AGL45424.1; provides kanamicin 
                     resistance in Escherichia coli"
                     /protein_id="AHE74077.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(2502..2593)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2609..3358)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     protein_bind    3516..3537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(3550..4632)
                     /codon_start=1
                     /gene="LacI"
                     /product="LacI"
                     /label=LacI
                     /note="lac repressor; derived from INSD CAA07594.1"
                     /protein_id="AHE74078.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(3550..4632)
                     /gene="LacI"
                     /label=LacI
     promoter        complement(4633..4710)
                     /gene="lacI"
                     /label=lacI promoter