LOCUS       Exported                3297 bp ds-DNA    circular SYN 25-11-2013
DEFINITION  Vector for high-efficiency cloning of DNA inserts with sticky or 
            blunt ends. A toxic lacZ-alpha-ccdB gene selects against clones 
            lacking an insert.
ACCESSION   .
VERSION     .
KEYWORDS    pZErO-2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3297)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3297
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     promoter        143..173
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             217..558
                     /codon_start=1
                     /gene="lacZ (fragment)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /note="lacZ-alpha"
                     /translation="MTMITPSYLGDTIEYSSYASSLVPSSDPLVTAASVLEFCRYPSHW
                     RPLEHASRGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSL"
     promoter        239..257
                     /note="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    269..381
                     /note="MCS"
                     /note="multiple cloning site"
     promoter        complement(389..407)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(414..430)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             568..870
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /note="ccdB"
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagaged in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     rep_origin      895..1274
                     /direction=RIGHT
                     /note="M13 ori"
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1322..2116)
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      2528..3116
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"