LOCUS       Exported                4433 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Gateway compatible binary vector pCG181, complete sequence.
ACCESSION   KT163016
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4433)
  AUTHORS   Wang G, Jones H, Doherty A, Yu X, Ma Y, Sun Y, Du W, Wang C, Xia L.
  TITLE     Generation of marker- and backbone-free transgenic wheat plants via 
            Agrobacterium-mediated transformation based on the pCLEAN dual 
            binary vector system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4433)
  AUTHORS   Wang G, Jones H, Doherty A, Yu X, Ma Y, Sun Y, Du W, Wang C, Xia L.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JUN-2015) Institute of Crop Sciences, Chinese Academy 
            of Agricultural Sciences, 12 Zhongguancun Street, Beijing, Beijing 
            100081, China
REFERENCE   3  (bases 1 to 4433)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4433
                     /organism="Gateway compatible binary vector pCG181"
                     /mol_type="other DNA"
                     /db_xref="taxon:1739641"
     misc_feature    1..2481
                     /note="Cloning vector pGreenII0000 (INSD EF590266) based 
                     backbone sequence"
     rep_origin      1..720
                     /label=pUC-ori
                     /note="pUC-ori"
     rep_origin      complement(61..649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(820..1635)
                     /codon_start=1
                     /gene="nptI"
                     /product="NptI"
                     /label=nptI
                     /note="kanamycin resistance"
                     /protein_id="ALK43903.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(820..1635)
                     /gene="nptI"
                     /label=nptI
     rep_origin      1921..2473
                     /label=pSa-ori
                     /note="pSa-ori"
     rep_origin      1926..2361
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    2508..2532
                     /note="T-DNA left border No.1 sequence (Thole et al., 
                     (2007) Plant Physiol 145:1211-1219)"
     misc_feature    2533..2558
                     /note="spacer sequence (Thole et al., (2007) Plant Physiol 
                     145:1211-1219)"
     misc_feature    2559..2583
                     /note="T-DNA left border No.2 sequence (Thole et al., 
                     (2007) Plant Physiol 145:1211-1219)"
     misc_recomb     2599..2723
                     /note="attR2; gateway recombination site; derived from 
                     Escherichia coli"
     protein_bind    2599..2723
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2764..3069)
                     /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="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(2773..3069)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /note="cell cycle death B; cytotoxic protein derived from 
                     Escherichia coli"
                     /protein_id="ALK43904.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFW"
     gene            complement(2773..3069)
                     /gene="ccdB"
                     /label=ccdB
     CDS             complement(3411..4070)
                     /codon_start=1
                     /gene="cmR"
                     /product="CmR"
                     /label=cmR
                     /note="chloramphenicol resistance protein"
                     /protein_id="ALK43905.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(3411..4070)
                     /gene="cmR"
                     /label=cmR
     promoter        complement(4124..4154)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_recomb     4179..4303
                     /note="attR1; gateway recombination site; derived from 
                     Escherichia coli"
     protein_bind    complement(4179..4303)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    4351..4375
                     /note="T-DNA right border sequence (Thole et al., (2007) 
                     Plant Physiol 145:1211-1219)"
     misc_feature    4376..4399
                     /note="right border overdrive sequence (Thole et al., 
                     (2007) Plant Physiol 145:1211-1219)"