LOCUS       Exported                4789 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Donor vector pZErOTM7ZeoGFPattB DNA, complete sequence.
ACCESSION   AB713996
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4789)
  AUTHORS   Yonemura N, Tamura T, Uchino K, Kobayashi I, Tatematsu K, Iizuka T, 
            Sezutsu H, Muthulakshmi M, Nagaraju J, Kusakabe T.
  TITLE     PhiC31 integrase-mediated cassette exchange in silkworm embryos
  JOURNAL   Mol. Genet. Genomics 287 (9), 731-739 (2012)
  PUBMED    22842670
REFERENCE   2  (bases 1 to 4789)
  AUTHORS   Yonemura N, Kobayashi I, Uchino K, Sezutsu H, Tamura T, Kusakabe T.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-APR-2012) Contact:Naoyuki Yonemura National Institute 
            of Agrobiological Sciences (NIAS), MAFF; Ohwashi 1-2, Tsukuba 
            Science City 305-8634, Japan
REFERENCE   3  (bases 1 to 4789)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4789
                     /organism="Donor vector pZErOTM7ZeoGFPattB"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /label=synonym: Donor plasmid pZErOTM7ZeoGFPattB
                     /note="synonym: Donor plasmid pZErOTM7ZeoGFPattB"
                     /db_xref="taxon:1177635"
     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"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_recomb     301..335
                     /label=Streptomyces phage phiC31 attB sequence
                     /note="Streptomyces phage phiC31 attB sequence"
     protein_bind    301..334
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     regulatory      complement(439..560)
                     /regulatory_class="terminator"
                     /note="SV40"
     polyA_signal    439..560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(624..1001)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MDAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRD
                     DVTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWG
                     REFALRDPAGNCVHFVAEEQD"
     CDS             complement(624..995)
                     /codon_start=1
                     /gene="Zeo"
                     /product="Zeocin resistant protein"
                     /label=Zeo
                     /note="Zeocin region of GFP-Zeocin fusion gene"
                     /protein_id="BAM36531.1"
                     /translation="AKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDDV
                     TLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGRE
                     FALRDPAGNCVHFVAEEQD"
     gene            complement(624..995)
                     /gene="Zeo"
                     /label=Zeo
     CDS             complement(996..1700)
                     /codon_start=1
                     /gene="GFP"
                     /product="green fluorescence protein"
                     /label=GFP
                     /note="GFP region of GFP-Zeocin fusion gene"
                     /protein_id="BAM36532.1"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMD"
     gene            complement(996..1700)
                     /gene="GFP"
                     /label=GFP
     regulatory      complement(1716..1782)
                     /regulatory_class="promoter"
                     /note="EM7 pormoter"
     promoter        complement(1734..1781)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     misc_recomb     1823..1857
                     /label=Streptomyces phage phiC31 attB sequence
                     /note="Streptomyces phage phiC31 attB sequence"
     protein_bind    1823..1856
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     misc_feature    1863..280
                     /label=pZErO-2-derived sequence
                     /note="pZErO-2-derived sequence"
     promoter        complement(1881..1899)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1906..1922)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2060..2362
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     rep_origin      2387..2766
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(2814..3608)
                     /codon_start=1
                     /gene="KmR"
                     /product="kanamycin resistance protein"
                     /label=KmR
                     /protein_id="BAM36533.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(2814..3608)
                     /gene="KmR"
                     /label=KmR
     rep_origin      complement(3994..4667)
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="ColE1 origin"
     rep_origin      4020..4608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"