LOCUS       Exported                5443 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBlueLysis, complete sequence.
ACCESSION   AY796342
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5443)
  AUTHORS   Yun J, Kang S, Ryu S.
  TITLE     Development of a novel vector system for programmed cell lysis in 
            Escherichia coli in response to glucose exhaustion
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5443)
  AUTHORS   Yun J, Kang S, Ryu S.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-OCT-2004) School of Agricultural Biotechnology, Seoul 
            National Univ., 200-8105 CALS Seoul National Univ. Kwanak-gu, Seoul 
            151-742, South Korea
REFERENCE   3  (bases 1 to 5443)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5443
                     /organism="Cloning vector pBlueLysis"
                     /mol_type="other DNA"
                     /db_xref="taxon:303227"
     protein_bind    143..164
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        179..209
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    217..233
                     /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     241..257
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             253..576
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDLRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     CDS             259..576
                     /codon_start=1
                     /gene="lacZ"
                     /product="LacZ alpha fragment"
                     /label=lacZ
                     /note="alpha fragment beta-galactosidase; cloned from 
                     pUC19; BamHI site was substituted with BglII"
                     /protein_id="AAV85010.1"
                     /translation="MITPSLHACRSTLEDLRVPSSNSLAVVLQRRDWENPGVTQLNRLA
                     AHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDA
                     A"
     gene            259..576
                     /gene="lacZ"
                     /label=lacZ
     misc_feature    270..326
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(327..343)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(1008..1553)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     promoter        1665..1693
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1741..2931
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline resistant protein"
                     /label=tet
                     /protein_id="AAV85011.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            1741..2931
                     /gene="tet"
                     /label=tet
     regulatory      3991..4170
                     /regulatory_class="promoter"
     CDS             4171..4824
                     /codon_start=1
                     /gene="t"
                     /product="holin"
                     /label=t
                     /note="from T4 bacteriophage"
                     /protein_id="AAV85012.1"
                     /translation="MAPRISFSPSDILFGVLDRLFKDNATGKVLASRVAVVILLFIMAI
                     VWYRGDSFFEYYKQSKYETYSEIIEKERTARFESVALEQLQIVHISSEADFSAVYSFRP
                     KNLNYFVDIIAYEGKLPSTISEKSLGGYPVDKTMDEYTVHLNGRHYYSNSKFAFLPTKK
                     PTPEINYMYSCPYFNLDNIYAGTITMYWYRNDHISNDRLESICAQAARILGRAK"
     gene            4171..4824
                     /gene="t"
                     /label=t
     CDS             4834..4851
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4958..5413
                     /codon_start=1
                     /gene="lys"
                     /product="lysozyme"
                     /label=lys
                     /note="from T7 bacteriophage"
                     /protein_id="AAV85013.1"
                     /translation="MARVQFKQRESTDAIFVHCSATKPSQNVGVREIRQWHKEQGWLDV
                     GYHFIIKRDGTVEAGRDEMAVGSHAKGYNHNSIGVCLVGGIDDKGKFDANFTPAQMQSL
                     RSLLVTLLAKYEGAGLRAHHEVAPKACPSFDLKRWWEKNELVTSDRG"
     gene            4958..5413
                     /gene="lys"
                     /label=lys
     promoter        5411..5443
                     /label=T7 Phi-3.8 promoter
                     /note="weak promoter for bacteriophage T7 RNA polymerase"