LOCUS       Exported                9272 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLZ2, complete sequence.
ACCESSION   KJ641600
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9272)
  AUTHORS   Zhu L, Dalbey RE.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAR-2014) Department of Chemistry and Biochemistry, 
            The Ohio State University, RM580, W.12th Ave, Columbus, OH 43210, 
            USA
REFERENCE   2  (bases 1 to 9272)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9272
                     /organism="Expression vector pLZ2"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /label=vector derived from pRSF-1b
                     /note="vector derived from pRSF-1b"
                     /db_xref="taxon:1509197"
     CDS             28..72
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     regulatory      110..171
                     /regulatory_class="terminator"
                     /note="T7"
     terminator      124..171
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(203..2854)
                     /codon_start=1
                     /product="T7 RNA polymerase"
                     /label=T7 RNA polymerase
                     /protein_id="AID07480.1"
                     /translation="MNTINIAKNDFSDIELAAIPFNTLADHYGERLAREQLALEHESYE
                     MGEARFRKMFERQLKAGEVADNAAAKPLITTLLPKMIARINDWFEEVKAKRGKRPTAFQ
                     FLQEIKPEAVAYITIKTTLACLTSADNTTVQAVASAIGRAIEDEARFGRIRDLEAKHFK
                     KNVEEQLNKRVGHVYKKAFMQVVEADMLSKGLLGGEAWSSWHKEDSIHVGVRCIEMLIE
                     STGMVSLHRQNAGVVGQDSETIELAPEYAEAIATRAGALAGISPMFQPCVVPPKPWTGI
                     TGGGYWANGRRPLALVRTHSKKALMRYEDVYMPEVYKAINIAQNTAWKINKKVLAVANV
                     ITKWKHCPVEDIPAIEREELPMKPEDIDMNPEALTAWKRAAAAVYRKDKARKSRRISLE
                     FMLEQANKFANHKAIWFPYNMDWRGRVYAVSMFNPQGNDMTKGLLTLAKGKPIGKEGYY
                     WLKIHGANCAGVDKVPFPERIKFIEENHENIMACAKSPLENTWWAEQDSPFCFLAFCFE
                     YAGVQHHGLSYNCSLPLAFDGSCSGIQHFSAMLRDEVGGRAVNLLPSETVQDIYGIVAK
                     KVNEILQADAINGTDNEVVTVTDENTGEISEKVKLGTKALAGQWLAYGVTRSVTKRSVM
                     TLAYGSKEFGFRQQVLEDTIQPAIDSGKGLMFTQPNQAAGYMAKLIWESVSVTVVAAVE
                     AMNWLKSAAKLLAAEVKDKKTGEILRKRCAVHWVTPDGFPVWQEYKKPIQTRLNLMFLG
                     QFRLQPTINTNKDSEIDAHKQESGIAPNFVHSQDGSHLRKTVVWAHEKYGIESFALIHD
                     SFGTIPADAANLFKAVRETMVDTYESCDVLADFYDQFADQLHESQLDKMPALPAKGNLN
                     LRDILESDFAFA"
     regulatory      complement(3226..3236)
                     /regulatory_class="promoter"
                     /note="pL; induces expression of T7 RNA polymerase"
     regulatory      complement(3247..3252)
                     /regulatory_class="minus_10_signal"
                     /note="lambda left promoter"
     regulatory      complement(3270..3275)
                     /regulatory_class="minus_35_signal"
                     /note="lambda left promoter"
     promoter        3415..3445
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    3453..3469
                     /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             3490..4203
                     /codon_start=1
                     /product="cI857"
                     /label=cI857
                     /note="lambda cI repressor protein"
                     /protein_id="AID07481.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQHPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             complement(4653..5468)
                     /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="AID07477.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(5469..5560)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5576..6325
                     /note="RSF origin; copy number above 100 per cell"
     CDS             complement(6521..7603)
                     /codon_start=1
                     /product="LacI"
                     /label=LacI
                     /note="lac repressor; derived from INSD CAA07594.1"
                     /protein_id="AID07478.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        7698..7716
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      7698..7714
                     /regulatory_class="promoter"
                     /note="T7"
     regulatory      7724..8014
                     /regulatory_class="promoter"
                     /note="lac promoter-operator"
     protein_bind    7781..7802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7817..7847
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7855..7871
                     /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)."
     primer_bind     7879..7895
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7911..7927)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      8007..8013
                     /regulatory_class="ribosome_binding_site"
     CDS             8021..2
                     /codon_start=1
                     /product="LacY"
                     /label=LacY
                     /note="lactose permease"
                     /protein_id="AID07479.1"
                     /translation="MYYLKNTNFWMFGLFFFFYFFIMGAYFPFFPIWLHDINHISKSDT
                     GIIFAAISLFSLLFQPLFGLLSDKLGLRKYLLWIITGMLVMFAPFFIFIFGPLLQYNIL
                     VGSIVGGIYLGFSFNAGAPAVEAFIEKVSRRSNFEFGRARMFGSVGWALVASIVGIMFT
                     INNQFVFWLGSGSALILAVLLFFAKTDAPSSATVANAVGANHSAFSLKLALELFRQPKL
                     WFLSLYVIGVSSTYDVFDQQFANFFTSFFATGEQGTRVFGYVTTMGELLNASIMFFAPL
                     IINRIGGKNALLLAGTIMSVRIIGSSFATSALEVVILKTLHMFEVPFLLVGSFKYITSQ
                     FEVRFSATIYLVSFSFFKQLAMIFMSVLAGNMYESIGFQGAYLVLGLVALGFTLISVFT
                     LSGPGPLSLLRRQVNEVA"