LOCUS       Exported               12907 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Gateway adapted binary vector pCGTAG, complete sequence.
ACCESSION   DQ370422
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12907)
  AUTHORS   Zhang C, Gong FC, Lambert GM, Galbraith DW.
  TITLE     Cell type-specific characterization of nuclear DNA contents within 
            complex tissues and organs
  JOURNAL   Plant Methods 1 (1), 7 (2005)
  PUBMED    16270943
REFERENCE   2  (bases 1 to 12907)
  AUTHORS   Zhang C, Galbraith DW.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JAN-2006) Department of Plant Sciences, University of 
            Arizona, 303 Forbes Building, Tucson, AZ 85721-0036, USA
REFERENCE   3  (bases 1 to 12907)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12907
                     /organism="Gateway adapted binary vector pCGTAG"
                     /mol_type="other DNA"
                     /db_xref="taxon:370581"
     misc_feature    1..1714
                     /label=Invitrogen attR cassette (reading frame C.1)
                     /note="Invitrogen attR cassette (reading frame C.1)"
     protein_bind    6..130
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        155..185
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             239..919
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             1239..1544
                     /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
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(1585..1709)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    1715..12907
                     /label=derived from plasmid pCsGFPBT
                     /note="derived from plasmid pCsGFPBT"
     5UTR           1715..1850
                     /note="translation leader sequence; 5UTR from tobacco etch
                     virus"
     CDS             1851..3032
                     /codon_start=1
                     /product="HTA6-sGFP"
                     /label=HTA6-sGFP
                     /note="translational fusion between HTA6 (Arabidopsis 
                     histone 2A, At5g59870) and sGFP (modified by the Sheen Lab 
                     with enhanced fluorescence)"
                     /protein_id="ABD16410.1"
                     /translation="MESTGKVKKAFGGRKPPGAPKTKSVSKSMKAGLQFPVGRITRFLK
                     KGRYAQRLGGGAPVYMAAVLEYLAAEVLELAGNAARDNKKSRIIPRHLLLAIRNDEELG
                     KLLSGVTIAHGGVLPNINSVLLPKKSATKPAEEKATKSPVKSPKKAAMVSKGEELFTGV
                     VPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFTYGVQC
                     FSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGID
                     FKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPI
                     GDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKGIL"
     CDS             2304..3020
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     misc_feature    3034..3103
                     /note="computer generated random sequence as a tag of the 
                     HTA6-sGFP fusion transcript"
     regulatory      3104..3319
                     /regulatory_class="polyA_signal_sequence"
                     /note="cauliflower mosaic virus 35S poly(A) signal"
     primer_bind     complement(3341..3357)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3950..4202
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      3953..4205
                     /regulatory_class="polyA_signal_sequence"
                     /note="nopaline synthase poly(A) signal"
     misc_feature    4224..4248
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     repeat_region   4243..4268
                     /label=right border T-DNA repeat
                     /note="right border T-DNA repeat"
     misc_feature    complement(5309..6309)
                     /label=STA region from pVS1 plasmid
                     /note="STA region from pVS1 plasmid"
     CDS             5548..6177
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6611..7678
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      complement(6902..7902)
                     /direction=LEFT
                     /note="pVS1-REP; replication origin from pVS1"
     rep_origin      7744..7938
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    8282..8422
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    complement(8312..8572)
                     /label=bom site from pBR322
                     /note="bom site from pBR322"
     rep_origin      complement(8608..9196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(8712..8992)
                     /direction=LEFT
                     /label=pBR322 replication origin
                     /note="pBR322 replication origin"
     CDS             complement(9283..10077)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=aminoglycoside phosphotransferase
                     /note="aadA (kanamycin resistance) amplified from pIG121Hm"
                     /protein_id="ABD16411.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     repeat_region   10502..10527
                     /label=left border repeat from C58 T-DNA
                     /note="left border repeat from C58 T-DNA"
     misc_feature    10502..10526
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     regulatory      complement(10594..10802)
                     /regulatory_class="polyA_signal_sequence"
                     /note="cauliflower mosaic virus 35S poly(A) signal"
     polyA_signal    10604..10778
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(10818..11843)
                     /codon_start=1
                     /product="hygromycin phosphotransferase"
                     /label=hygromycin phosphotransferase
                     /note="hptII (hygromycin resistance)"
                     /protein_id="ABD16412.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     regulatory      complement(11879..12659)
                     /regulatory_class="promoter"
                     /note="CaMV35S2; CaMV 35S promoter; duplicated"
     promoter        complement(11911..12588)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    12779..12800
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      12807..12861
                     /regulatory_class="promoter"
                     /note="lacZ promoter"
     promoter        12815..12845
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12853..12869
                     /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     12877..12893
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"