LOCUS       Exported                3356 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pZero++ Kan, complete sequence.
ACCESSION   AY569777
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3356)
  AUTHORS   Wu G, Zhu Z.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAR-2004) Fish Genetics, Institute of Hydrobiology, 
            Chinese Academy of Science, 7# South Road of East Lake, Wuhan, Hubei
            430072, Peoples Republic of China
REFERENCE   2  (bases 1 to 3356)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3356
                     /organism="Cloning vector pZero++ Kan"
                     /mol_type="other DNA"
                     /db_xref="taxon:268731"
     regulatory      95..216
                     /regulatory_class="promoter"
                     /note="Lac promoter/operator"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             217..930
                     /codon_start=1
                     /product="LacZ alpha & ccdB lethal fusion protein"
                     /label=LacZ alpha & ccdB lethal fusion protein
                     /protein_id="AAS78491.1"
                     /translation="MTMITPSYLGDTIEYSSYASDLICLGAPRTSTSLVPSMTWRHGSL
                     GDPLVTAASVLEFCRYPSHWRPLEHASRGPNSPYSESYYNSLAVVLQRRDWENPGVTQL
                     NRLAAHPPFASWRNSEEARTDRPSQQLRSLYVRQFKVYTYKRESRYRLFVDVQSDIIDT
                     PGRRMVIPLASARLLSDKVSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHR
                     ENDIKNAINLMFWGI"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        complement(449..467)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(474..490)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      955..1367
                     /label=f1
                     /note="f1"
     rep_origin      955..1334
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1382..2176)
                     /codon_start=1
                     /product="Kan"
                     /label=Kan
                     /protein_id="AAS78490.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      2561..3234
                     /label=Col E1
                     /note="Col E1"
     rep_origin      2587..3175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"